Soluble polypeptide fractions of the LAG-3 protein, production method, therapeutic composition, anti-idiotype antibodies

ABSTRACT

Soluble polypeptide fraction consisting of all or part one at least of the four immunoglobulin-type extracellular LAG-3 protein domains (amino acids 1-159, 160-230, 240-330 and 331-412 of the SEQ ID NO:1 sequence) or consisting of one peptide sequence derived from these domains by replacement, addition or deletion of one or more amino acids. The fraction of the invention has a specificity at least equal to that of LAG-3 in relation to its ligand.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a §371 application of PCT/FR95/00593, filed May 5, 1995.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to soluble forms derived from the LAG-3 membrane protein which are useful as immunosuppressants, as well as antibodies capable of preventing the specific binding of the LAG-3 protein to MHC (major histocompatibility complex) Class II molecules as immunostimulants.

2. Description of the Related Art

In WO-A 91/10682, a protein designated LAG-3 has been described.

The LAG-3 protein is a protein selectively expressed by NK cells and activated T lymphocytes. Similarity of the amino acid sequence, the comparative exon/intron organization and the chromosomal localization show that LAG-3 is related to CD4. The initial characterization of the LAG-3 gene has been described by TRIEBEL et al. (1).

The corresponding DNA codes for a type I transmembrane protein of 498 amino acids containing 4 extra-cellular sequences of the immunoglobulin type. LAG-3 is a member of the immunoglobulin superfamily.

The mature protein comprises 476 amino acids (SEQ ID No. 1) with a theoretical molecular weight of 52 kD. The extracellular region contains 8 cysteine residues and 4 potential N-glycosylation sites. By Western blot analysis, it was shown that LAG-3 inside PRA-blasts or activated NK cells has an apparent mass Mr of 70,000. After treatment with N-glycosidase F, a reduction in size to 60 kD was obtained, thereby demonstrating that native LAG-3 is glycosylated. Fuller details are described in WO-A 91/10682.

BAIXERAS et al., in J. Exp. Med. 176, 327-337 (2), have, in addition, described their finding that rosette formation between cells transfected with LAG-3 (expressing LAG-3 at their surface) and B lymphocytes expressing MHC Class II was specifically dependent on LAG-3/MHC Class II interaction.

Surprisingly, this ligand for MHC Class II was detected with higher levels on activated CD8⁺ lymphocytes (MHC Class I-restricted) than on activated CD4⁺ lymphocytes. In vivo, only a few disseminated LAG-3⁺ cells (MHC Class II-restricted) were to be found in non-hyperplastic lymphoid tissue comprising the primary lymphoid organs, that is to say thymus and bone marrow. LAG-3⁺ cells were to be found in hyperplastic lymphoid nodules and tonsils, as well as among peripheral blood mononuclear cells (PBMC) of patients receiving injections of high doses of IL-2.

These observations confirm that LAG-3 is an activation antigen in contrast to CD4 expressed in a subpopulation of resting lymphocytes and other cell types, in particular macrophages.

The MHC comprises Class I and Class II molecules which are membrane glycoproteins which present fragments of protein antigens to the T lymphocyte receptors (TCR). Class I molecules are responsible for the presentation to CD8⁺ cytotoxic cells of peptides derived in large part from endogenously synthesized proteins, while Class II molecules present to CD4⁺ helper lymphocytes peptides originating in the first place from foreign proteins which have entered the endocytic, that is to say exogenous, pathway. T helper lymphocytes regulate and amplify the immune response, while cytotoxic lymphocytes are needed to destroy cells irrespective of the tissues expressing "non-self" antigens, for example viral antigens. The mechanism of recognition involves intracellular signals leading to an effective activity of T lymphocytes.

It is apparent that, to initiate an immune response mediated by T (CD4⁺) lymphocytes, the foreign antigens must be captured and internalized in the form of peptides by specialized cells, the antigen presenting cells (APC). The resulting antigenic peptides are reexpressed at the surface of the antigen presenting cells, where they are combined with MHC Class II molecules. This MHC Class I/peptide complex is specifically recognized by the T lymphocyte receptor, resulting in an activation of the T helper lymphocytes.

Moreover, animal models created by recombination techniques have made it possible to emphasize the part played in vivo by MHC Class II molecules and their ligands.

Thus, mice deficient in MHC Class II molecules (3) and possessing almost no peripheral CD4⁺ T lymphocytes and having only a few immature CD4⁺ lymphocytes in the thymus have proved to be completely incapable of responding to T-dependent antigens.

CD4⁻ /⁻ mutant mice (4) have a substantially decreased T lymphocyte activity but show normal development and function of the CD8⁺ T lymphocytes, demonstrating that the expression of CD4 on the daughter cells and CD4⁺ CD8⁺ thymocytes is not obligatory for the development. Compared to normal mice, these CD4-deficient mice have a large amount of CD4⁻ CD8⁻ cells.

These doubly negative cells are restricted to MHC Class II and capable of recognizing the antigen.

When they are infected with Leishmania, these mice show a population of functional T helper lymphocytes despite the absence of CD4. These cells are restrictive to MHC Class II and produce interferon-γ when they are activated by the antigen. This indicates that the lineage of the T lymphocytes and their peripheral function need not necessarily depend on the function of CD4.

It is now recognized that the proteins encoded by MHC Class II region are involved in many aspects of immune recognition, including the interaction between different lymphoid cells such as lymphocytes and antigen presenting cells. Different observations have also shown that other mechanisms which do not take place via CD4 participate in the effector function of T helper lymphocytes.

These different observations underline the pivotal role played by MHC Class II and its ligands in the immune system.

Moreover, the importance is known of chimeric molecules composed of the extracytoplasmic domain of proteins capable of binding to ligands and a constant region of human immunoglobulin (Ig) chains for obtaining soluble forms of proteins and of cell receptors which are useful, in particular, as therapeutic agents.

Thus, soluble forms of CD4 have proven their efficacy in inhibiting an HIV infection in vitro in a dose-dependent manner.

Nevertheless, clinical trials with soluble CD4 molecules, in particular of CD4-Ig, have not enabled a significant decrease in viral titres to be demonstrated. Transgenic mice expressing up to 20 μg/ml of soluble CD4 in their serum were created. These mice showed no difference as regards their immune function relative to control mice. Hitherto, no direct binding to MHC Class II of molecules derived from CD4 has been reported. This strongly suggests that soluble CD4 molecules do not interact in vivo with MHC Class II molecules.

SUMMARY OF THE INVENTION

Surprisingly, the authors of the present invention have shown that soluble molecules containing different fragments of the extracytoplasmic domain of the LAG-3 protein were capable of binding to MHC Class II molecules and of having an immunosuppressant action.

The extracytoplasmic region of LAG-3 represented by the sequence SEQ ID No. 1 comprises the domains D1, D2, D3 and D4 extending from amino acids 1 to 159, 160 to 239, 240 to 330 and 331 to 412, respectively.

Thus, the subject of the invention is a soluble polypeptide fraction consisting of all or part of at least one of the 4 immunoglobulin type extracellular domains of the LAG-3 protein (amino acid 1 to 159, 160 to 239, 240 to 330 and 331 to 412 of the sequence SEQ ID No. 1), or of a peptide sequence derived from these domains by replacement, addition and/or deletion of one or more amino acids, and which possesses a specificity at least equal to or greater than that of LAG-3 for its ligand.

The present invention encompasses, in particular, soluble polypeptide fractions having a sequence derived from the native LAG-3 sequence originating from the well-known phenomenon of polytypy.

The soluble polypeptide fraction is characterized in that it comprises the peptide region of LAG-3 responsible for the affinity of LAG-3 for MHC Class II molecules.

The soluble polypeptide fraction comprises, in particular, a peptide sequence derived from these domains by replacement, addition and/or deletion of one or more amino acids, and which possesses a specificity equal to or greater than that of LAG-3 for its ligand, for example the whole of the first two immunoglobulin type domains of LAG-3, or the 4 immunoglobulin type domains of the extracytoplasmic domain of LAG-3.

Advantageously, the soluble polypeptide fraction is comprised of all or part of at least one of the four immunoglobulin type extracellular domains of the LAG-3 protein (amino acid 1 to 149, 150 to 239, 240 to 330 and 331 to 412 of sequence SEQ ID No. 1) comprising one or more of the arginine (Arg) rests at the positions 73, 75 and 76 of sequence SEQ ID No. 1 substituted with glutamic acid (Glu).

Preferably, the soluble polypeptide fraction comprises a loop in which the average position of the atoms forming the basic linkage arrangement is given by the position of amino acids 46 to 77 (SEQ ID No. 1) appearing in Table 1 or Table 2 or differs therefrom by not more than 5%.

The soluble polypeptide fraction advantageously comprises, in addition, the second immunoglobulin type extracellular domain (D2) of LAG-3 (amino acids 150 to 241).

Advantageously, the soluble polypeptide fraction comprises, besides the peptide sequence of LAG-3 as defined above, a supplementary peptide sequence at its C-terminal and/or N-terminal end, so as to constitute a fusion protein. The term "fusion protein" means a portion of any protein permitting modification of the physicochemical features of the subfragments of the extracytoplasmic domain of the LAG-3 protein. Examples of such fusion proteins contain fragments of the extracytoplasmic domain of LAG-3 as are defined above, bound to the heavy chain --CH2--CH3 junction region of a human immunoglobulin, preferably an isotype IgG4 immunoglobulin.

Such fusion proteins may be dimeric or monomeric. These fusion proteins may be obtained by recombination techniques well known to a person skilled in the art, for example a technique such as that described by Traunecker et al. (5).

Generally speaking, the method of production of these fusion proteins comprising an immunoglobulin region fused with a peptide sequence of LAG-3 as defined above consists in inserting into a vector the fragments of cDNA coding for the polypeptide regions corresponding to LAG-3 or derived from LAG-3, where appropriate after amplification by PCR, and the cDNA coding for the relevant region of the immunoglobulin, this cDNA being fused with cDNA coding for the corresponding polypeptide regions or derivatives of LAG-3, and in expressing after transfection the fragments cDNA in an expression system, in particular mammalian cells, for example hamster ovary cells.

The fusion proteins according to the invention may also be obtained by cleavage of a LAG-3/ Ig conjugate constructed so as to contain a suitable cleavage site.

The subject of the invention is also a therapeutic composition having immunosuppressant activity comprising a soluble polypeptide fraction according to the invention. This composition will be useful for treating pathologies requiring immunosuppression, for example autoimmune diseases.

The subject of the invention is also the use of antibodies directed against LAG-3 or soluble polypeptide fractions derived from LAG-3 as are defined above, or fragments of such antibodies, in particular the Fab, Fab' and F(ab')₂ fragments, for the preparation of a therapeutic composition having immunostimulatory activity. "Immunostimulatory" means a molecular entity capable of stimulating the maturation, differentiation, proliferation and/or function of cells expressing LAG-3, that is to say T lymphocytes or active NK cells. The anti-LAG-3 antibodies may be used as potentiators of vaccines or immunostimulants in immunosuppressed patients, such as patients infected with HIV or treated with immunosuppressant substances, or be used to stimulate the immune system by elimination of self cells displaying abnormal behaviour, for example cancer cells.

Immunostimulatory activity of anti-LAG-3 anti-bodies is surprising, inasmuch as anti-CD4 antibodies have an immunosuppressant action.

Such antibodies may be polyclonal or monoclonal; however, monoclonal antibodies are preferred. The polyclonal antibodies may be prepared according to well-known methods, such as that described by BENEDICT A.A. et al. (6). Monoclonal antibodies are preferred, on account of the fact that they are specific for a single epitope and yield results with better reproducibility. Methods of production of monoclonal antibodies are well known from the prior art, especially the one described by KOHLER and MILSTEIN. This method, together with variants thereof, are described by YELTON et al. (7).

The subject of the invention is also anti-idiotype antibodies directed against the antibodies according to the invention, which contain the internal image of LAG-3 and are consequently capable of binding to MHC Class II. Such antibodies may be used, in particular, as immunosuppressants, and, for example, in autoimmune pathologies.

The therapeutic compositions according to the present invention comprise soluble LAG-3 proteins or antibodies as are defined above, as well as a pharmaceutically acceptable vehicle. These compositions may be formulated according to the usual techniques. The vehicle can vary in form in accordance with the chosen administration route: oral, parenteral, sublingual, rectal or nasal.

For the compositions for parenteral administration, the vehicle will generally comprise sterile water as well as other possible ingredients promoting the solubility of the composition or its ability to be stored. The parenteral administration routes can consist of intravenous, intramuscular or subcutaneous injections.

The therapeutic composition can be of the sustained-release type, in particular for long-term treatments, for example in autoimmune diseases. The dose to be administered depends on the subject to be treated, in particular on the capacity of his/her immune system to achieve the desired degree of protection. The precise amounts of active ingredient to be administered may be readily determined by the practitioner who will initiate the treatment.

The therapeutic .compositions according to the invention can comprise, in addition to soluble LAG-3 or the antibodies according to the invention, another active ingredient, where apprto LAG-3 or to anmical bond to LAG-3 or to an antibody according to the invention. As an example, there may be mentioned soluble LAG-3 proteins according to the invention fused to a toxin, for example ricin or diphtheria toxoid, capable of binding to MHC Class II molecules and of killing the target cells, for example leukaemic or melanoma cells, or fused to a radioisotope.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a comparison of the proliferation of T cells incubated with F(ab) fragments of 17B4 to the proliferation of T cells incubated with intact 17B4 monoclonal antibody.

FIG. 2 shows the proliferation of Clone 28 in response to tetanus toxoid when co-cultured with 17B4 or control antibody 10H3.

FIG. 3 shows the expression vector pCDM7 used for manufacturing the recombinant LAG-3 proteins and a recombinant CD8 immunoadhesin control.

FIG. 4 shows the pCLH3 AXS V2 DHFR hα IVS vector used to express amplified LAG-3 sequences.

FIG. 5A shows the inhibition of MHC Class II interaction with LAG-3 by recombinant LAG-3 D1-D4 and FIG. 5B shows the potential inhibition of MHC Class II interaction with CD4.

FIG. 6 shows the inhibition of Clone 28 proliferation by recombinant LAG-3 D1-D4.

FIG. 7 shows the inhibition of Clone 154 proliferation by LAG-3 Ig.

FIG. 8 shows the binding of LAG-3 to B cell lines expressing MHC class II haplotypes or human class II-transfected mouse cells.

FIG. 9 shows the binding of LAG-3 Ig to MHC Class II expressing Daudi cells.

FIG. 10 shows that preincubation of HLA class II expressing cells with 17B4 inhibits LAG-3 Ig binding.

FIGS. 11A-11C show the inhibition of clone T154 proliferation by crosslinked LAG-3 Ig.

FIG. 12 compares inhibition of T cell proliferation by anti-Class II antibodies to inhibition of T cell proliferation by LAG-3 Ig.

FIGS. 13A-13D show that T cell proliferation in response to OKT3 (FIG. 13A), lectins (FIG. 13B) and low-concentration IL₂ (FIG. 13C) is inhibited by LAG-3 Ig but proliferation in response to high-concentration IL₂ is not inhibited by LAG-3 Ig (FIG. 13D).

FIG. 14 shows clone S1B5 cytotoxicity towards Epstein-Barr virus transformed human B cells.

FIG. 15 shows peripheral blood lymphocyte cytotoxicity towards HLA Class I⁻ Daudi cells.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The examples which follow, together with the attached reference figures, will illustrate the invention in greater detail.

EXAMPLE 1 Proliferation of active T lvmphocvte lines in the presence of anti-LAG-3 monoclonal antibodies

The anti-LAG-3 monoclonal antibodies used were 17B4, described in BAIXERAS et al. (2) and deposited at the CNCM under No. I-1240 on Jul. 10, 1992, and 11E3, described in HUARD et al. (8).

These antibodies belong to the isotype IgG1. These antibodies were tested for their biological effects on activated T lymphocytes, stimulated by specific antigenic peptides or processed antigens presented by MHC Class II molecules expressed by autologous antigen presenting cells, expressing LAG-3.

An anti-CD48 monoclonal antibody designated 10 H3 was used as irrelevant IgG1 antibody (negative control).

The saturating concentrations of anti-LAG-3 and anti-CD48 antibodies were determined by immunofluorescence on PHA (phytohaemagglutinin)-blasts and cell lines transformed by Epstein-Barr virus (EBV). In the proliferation tests, the monoclonal antibodies were added in the proportion of 5 times the saturating concentration.

The T lymphocyte lines used were, on the one hand the clone 154 derived from peripheral blood lymphocytes, raised against a peptide mimicking an influenza haemagglutinin (HA) fragment having an amino acid sequence extending from amino acid 306 to 329 (p20 peptide), and on the other hand the clone 28, a T lymphocyte clone derived from peripheral lymphocytes of a single human donor, raised against diphtheria toxoid (DT). The antigen presenting cells (APC) corresponding to clone 154 were EBV-transformed B lymphocytes of the same donor (DR3/DR11) as T 154. The antigen presenting cells corresponding to clone 28 were EBV-transformed B lymphocytes of the same donor. This clone was restricted to HLA DR7.

For clone 154, the APC (5×10⁶) were incubated at 37° C. for one and a half hours with variable doses of the p20 peptide, then washed and irradiated (10,000 rad). The cells were plated out on 96-well microtitration plates at the same time as the clone 154 cells (0.5×10⁵ to 10×10⁵ cells/ml) in a 3:1 ratio. For clone 28, the responding cells/stimulating cells ratio was 1.

The HLA DR7/EBV APC cells were either treated with mitomycin or irradiated, then added to the T lymphocytes in the presence of DT (which remained in the culture). The final concentration of clone 28 cells was 100,000 cells/ml.

³ H!Thymidine (1 μCi/well) was added at varying time intervals from day 2 to day 10 of culture.

Each experiment was carried out in triplicate.

The results were expressed as the mean cpm and after subtraction of the cpm found in the negative control (T lymphocytes cocultured with APC unladen with immunogens). The proliferation tests were carried out on 96-well plates. The absorption of tritiated thymidine in the individual 200 μl wells was measured after adding 1 μCi of thymidine for the last 18 hours of culture. The results were expressed in the form of the mean of 3 tests. The standard deviation was usually less than 12% (a little more in the case of very low cpm measurements). Moreover, mixed culture (clone 154/APC) supernatants were combined, filtered through 0.22 μm membranes, divided into samples and frozen at -20° C. until the time of titration using commercial immunoassay kits: Immunotech IL-2 and INF-α titration kit, Genzyme IFN-γ kit and Cayman Chemicals IL-4 kit.

A dose determination study was carried out to establish the proliferation profiles of clone 154 brought into contact with the p20 specific antigen at varying concentrations and in the presence or absence of anti-LAG-3 monoclonal antibodies or irrelevant monoclonal antibodies (negative control).

The individual results of 16 separate tests showed that, irrespective of the concentration of added antigen, the initial point up to the peak of proliferation was not modified, but a significant prolongation of the proliferation of T lymphocytes incubated with the anti-LAG-3 monoclonal antibodies was observed systematically. Fab fragments of the monoclonal antibody 17B4 were prepared and used in a test of proliferation of clone 154. The proliferation profile of T lymphocytes activated by the antigen with the 17B4 Fab fragments (15 μg/ml) was similar to that of cells incubated in the presence of whole 17B4 monoclonal antibody (40 μg/ml) (FIG. 1). These results show that the observed biological effects are not attributable to a non-specific reaction induced by the Fc region of the anti-LAG-3 monoclonal antibodies.

Similar results were obtained with the 11E3 anti-LAG-3 monoclonal antibodies.

Clone 28 was also stimulated with the antigen (tetanus toxoid 10 μg/ml) in the presence of 17B4 monoclonal antibodies after coculture with the corresponding APC in the presence of DT. The results are shown in FIG. 2.

The effects of the anti-LAG-3 monoclonal antibodies observed with clone 28, namely the prolongation of proliferation, are similar to those observed with clone 154.

Tests were carried out designed to measure the miscellaneous cellular events occurring after the antigenic stimulation of clone 154 cells incubated in the presence of anti-LAG-3 monoclonal antibodies.

The cells were harvested during conventional antigenic stimulation of clone 154 in the presence of anti-LAG-3 or anti-CD48 monoclonal antibodies or in the absence of antibodies, and tested for the expression of LAG-3 and CD25 transmembrane receptors, and samples of culture supernatants were collected at different time intervals after stimulation and tested for the presence of IFN-γ, TNF-α, IL-4 and IL-2.

Two-colour direct immunofluorescence tests (anti-CD3 monoclonal antibodies and anti-CD25 monoclonal antibodies) showed that IL-2 receptors were weakly but significantly increased 5 days after the antigenic stimulation. Similar tests with anti-CD3 and 11E3 (anti-LAG-3) monoclonal antibodies showed that LAG-3 was over-expressed from the day following activation onwards. In addition, the secretion of IL-2, IL-4, IFN-γ and TNF-α was also modulated by incubation with anti-LAG-3 monoclonal antibodies, thus showing that different cellular events are modified by the presence of anti-LAG-3 monoclonal antibodies and that some events already take place 24 hours after stimulation.

These results show indirectly that LAG-3 plays a regulatory role for CD4⁺ cells. The fact that anti-LAG-3 monoclonal antibodies increase proliferation, and hence act as immunopotentiators, suggest that LAG-3 is involved in the "deactivation" of CD4⁺ T lymphocytes with a negative role of LAG-3 on the antigen-dependent stimulation.

EXAMPLE 2 Transient expression of LAG-3 fusion proteins

Soluble proteins derived from LAG-3 were obtained by a recombinant DNA technique using suitable vectors comprising DNA coding for LAG-3 and DNA coding for an immunoglobulin fragment. The transient expression system consisted of transfected Cos cells. This system makes it possible to produce several mg of recombinant fusion proteins. Recombinant DNA techniques were carried out as described by MANIATIS et al. (22). The modifications were made as recommended by the manufacturer.

Construction of LAG-3 D1-D4 Ig and LAG-3 D1D2 Ig

Fragments coding for the D1D2 or D1-D4 regions were amplified (30 cycles) from a fragment of cDNA (FDC sequence) encompassing LAG-3 cDNA (TRIEBEL et al. (1)), using Taq polymerase free from 5'-endonuclease activity and relatively resistant to an exposure to very high temperature; the amplification was followed by a denaturation at 98° C. (with a Perkin Elmer Cetus "DNA thermal cycle"). Specific primers were used as recorded in the table below.

The resulting amplified fragments (739 bp and 1312 bp for LAG-3 D1-D2 and LAG-3 D1-D4, respectively) were inserted into a pBS plasmid (Stratagene).

Inserts were prepared after digestion with XhoI and BglII and introduced into the XhoI/BamHI sites of the vector pCDM7-CD8-IgG1 (pCDM7 being derived from pCDM8 marketed by Stratagene), as illustrated in FIG. 3, so as to exchange the DNA sequences coding for CD8 for those coding for the subfragments of LAG-3. The resulting expression vectors contained the sequences coding for D1D2 or D1-D4 fused to the DNA sequences coding for the --CH₂ --CH₃ junction region of a human IgG1 chain.

                                      TABLE 3     __________________________________________________________________________     Primers used to amplify LAG-3 DNA sequences by PCR                                          Resulting                                          encoded subfrag-                                          ment fused with     Primers used for amplification of the DNA                                          a subfragment Ig     __________________________________________________________________________     Primer (5')                          LAG-3 D1D2     5' GCGCCTCGAGGCCCAGACCATAGGAGAGATGT 3' (SEQ ID NO: 2)                                          from the leader     coupling          untranslated                start of                  sequence to     site 5' sequences                translation               amino acid 241     Primer (3')     5' GCGCAGATCTCTCCAGACCCAGAACAGTGAGGTTATACAT 3' (SEQ ID NO: 3)     BglII coup-            End of D2     ling site     Primer (5')                          LAG-3 D1-D4     identical to LAG-3 D1D2              from the leader     Primer (3')                          sequence to     5' GCGCAGATCTACCTGGGCTAGACAGCTCTGTGAA 3' (SEQ ID NO: 4)                                          amino acid 412     Bg1II coup-            End of D4     ling site     __________________________________________________________________________

CDM7 is a eukaryotic expression vector derived from the vectors developed by SEED et al. (10) for the cloning of DNA and its expression in E. coli and eukaryotic cells. CDM7 possesses the following features: (i) the human cytomegalovirus promoter for transient expression in mammalian cells; (ii) a viral origin of SV40 for an autosomal replication of mammalian cells expressing T antigen; (iii) πVX (type Col E1) as plasmid origin for a high copy number; (iv) a Sup F selection for resistance to ampicillin and tetracycline in Tet^(amb) and Amp^(amb) E. coli strains; (v) an origin of replication of M13 for the release of a single strand; (vi) a T7 RNA promoter; and (vii) a polylinker for an efficient cloning of heterologous DNA.

Transient expression in Cos cells

Cos cells (5×10⁶) were transfected with 30 μg of DNA of suitable expression vectors (coding for either LAG-3 D1D2 Ig, or LAG-3 D1-D4 Ig, or CD8 Ig) by electroporation (200 V, 1500 μF, 30-40 msec) using a Cellject apparatus (Eurogentech, Liege, BE). The cells were plated out again and cultured on a medium containing 5% of foetal calf serum. The supernatants were withdrawn 6 days after transfection.

The synthesis of the resulting fusion proteins was analysed from the supernatants as well as from cell extracts of transfected cells, by Western blot analysis with the 17B4 monoclonal antibodies. Immunoreactive materials were observed in the supernatant of cells transfected with DNA coding for LAG-3 D1D2 Ig or LAG-3 D1-D4 Ig.

Concomitantly, a recombinant CD8 immunoadhesin (CD8 Ig) was obtained as negative control using the same expression system and the expression vector pCDM7-CD8 (FIG. 3).

The recombinant proteins LAG-3 D1D2 Ig, LAG-3 D1-D4 Ig and CD8 Ig were purified by means of the standard method on protein A-Sepharose. The resulting material was analysed by SDS-PAGE, followed by Coomassie staining or a Western blot analysis using anti-human Ig antibody.

EXAMPLE 3 Production of soluble subfragments of LAG-3

In order to produce large amounts of recombinant proteins, a stable expression system consisting of transfected mammalian cells was developed. The host cells are anchorage-dependent hamster ovary (CHO) cells isolated from CHO cells deficient in dihydrofolate reductase (DHFR) and consequently necessitating glycine, a purine and thymidine for their growth. The pivotal role of DHFR in the synthesis of nucleic acid precursors, combined with the sensitivity of DHFR-deficient cells with respect to tetrahydrofolate analogues such as methotrexate (MTX), has two major advantages. Transfection of these cells with expression vectors containing the DHFR gene permits the secretion of recombinant DHFR-resistant clones, and the culturing of these cells on selective media containing increasing amounts of MTX results in amplification of the DHFR gene and the DNA associated therewith.

Construction of LAG-3 D1, LAG-3 D1D2, LAG-3 D1-D4

Fragments of DNA coding for the D1, D1D2 or D1-D4 regions were amplified using a PCR method identical to the one described previously, using the primers specified in the table below.

                                      TABLE 4     __________________________________________________________________________     Primers used for amplifying LAG-3 DNA sequences by PCR                                          Resulting                                          encoded     Primers used for amplification of the DNA                                          subfragment     __________________________________________________________________________     Primer (5')                          LAG-3 D1     5' CGCCGTCGACCGCTGCCCAGACCATAGGAGAGATGTG 3' (SEQ ID NO:                                          from the leader     SalI coup-          untranslated                start of                  sequence to     ling site          5' sequences                translation               amino acid 149     Primer (3')     5' GCGCGTCGACTTAACCCAGAACAGTGAGGTTATAC 3' (SEQ ID NO: 6)     SalI coup-            End of D1     ling site     Primer (5')                          LAG-3 D1D2     identical to LAG-3 D1                from the leader     Primer (3')                          sequence to     5' GCGCGTCGACTTAACCCAGAACAGTGAGGTTATAC 3' (SEQ ID NO: 7)                                          amino acid 239     SalIII coup-            End of D2     ling site     Primer (3')                          amino acid 149     5' GCGCGTCGACTTAACCCAGAACAGTGAGGTTATAC 3' (SEQ ID NO: 6)     SalI coup-            End of D1     ling site     Primer (5')                          LAG-3 D1-D4     identical to LAG-3 D1                from the leader     Primer (3')                          sequence to     5' GCGCGTCGACTTAACCCTGGGCTAGACAGCTCTCTGTG 3' (SEQ ID NO:                                          amino acid 412     SalI coup-            End of D4     ling site     __________________________________________________________________________

The resulting amplified fragments were digested with SalI and inserted into the SalI site of pUC 18 (Stratagene).

The amplified sequences were verified, and the inserts subcloned into the expression vector pCLH3 AXS V2 DHFR hα IVS as described by COLE et al. (Biotechnology 11, 1014-1024, 1993) (FIG. 4).

This vector is a eukaryotic expression vector which is multifunctional for the expression CDNA and its amplification in eukaryotic cells. It possesses the following features: (i) the murine promoter of the metallothionein-1 gene and a polyadenylation sequence SV 40 (comprising a donor-acceptor splicing site) to bring about transcription of the gene of interest, (ii) a human intervening sequence A containing the donor-acceptor splicing site of the gene for the subunit of α glycoprotein for obtaining high levels of transcription of cDNA, (iii) the pML sequence containing the origin of replication of pBR 322 and a gene for resistance to aampicillin for bacterial amplification, and (iv) a DHFR transcription unit of SV 40 to bring about transcription of the sequences used for selection and amplification of the transfectants.

Stable expression in CHO cells

The expression vectors coding for LAG-3 D1, LAG-3 D1D2 and LAG-3 D1-D4 were used to transfect CHO DUKX cells, and these cells were cultured on a selective medium. Cells capable of multiplying under these conditions were combined and cultured on a medium containing increasing amounts of MTX. Levels of expression were measured by Western blot analysis using the 17B4 monoclonal antibody. Clones producing high levels of recombinant soluble molecules derived from LAG-3 were propagated in bioreactors, and the material derived from LAG-3 was purified by ion exchange chromatography and immunoaffinity.

Western blot analyses revealed, in supernatants of cells transfected with expression vectors coding for LAG-3 D1, LAG-3 D1D2 and LAG-3 D1-D4, bands with apparent Mr values of 15 to 18 kD, 34-36 kD (doublets) and 55 kD (2 possible bands) . The respective Mr values of these immunoreactive materials corresponded to the expected Mr values of glycosylated LAG-3 D1 Ig (139 amino acids and a putative N-glycosylation site), glycosylated LAG-3 D1D2 Ig (239 amino acids containing 3 glycosylation sites) and glycosylated LAG-3 D1-D4 Ig (412 amino acids containing 4 glycosylation sites).

EXAMPLE 4 Specific binding of LAG-3 Ig to cells expressing MHC Class II

The reactivity of the monoclonal antibodies and of LAG-3 D1-D4 Ig was studied by indirect immunofluorescence. Target cells (4×10⁵) were incubated for 30 minutes at 4° C. in the presence of LAG-3 D1-D4 Ig, CD8 Ig, a murine monoclonal antibody, (949) anti-human MHC Class II (DR, DP, DQ) conjugated to FITC (isothiocyanate fluoride) from a Coulter clone, or murine Ig-FITC: an irrelevant immunoglobulin G conjugated to FITC. The cells were washed and incubated at 4° C. for 30 minutes with either a goat anti-human Ig polyclonal F(ab')₂ conjugated to fluorescein or a goat anti-mouse Ig polyclonal antibody conjugated to fluorescein (Coulter clone).

To confirm the LAG-3/MHC Class II binding, LAG-3 D1-D4 Ig was incubated with MHC Class II-positive or -negative cells. Four B lymphocyte lines expressing MHC Class II(L31, Phil EBV, Raji, Sanchez and Personnaz) were treated with anti-Class II monoclonal antibody 949, or the supernatants of Cos cells transfected with DNA coding either for LAG-3 D1-D4 Ig or for CD8 Ig. The five cell lines expressing the different haplotypes of MHC Class II molecules were recognized by LAG-3 Ig in the same way as by the anti-Class II monoclonal antibodies (positive control), while the supernatant containing CD8 Ig (negative control) did not bind to these cell lines, as could be expected. Four MHC Class II-negative cell lines (CEM, RJ, HSB2, K562) were treated with the same reagents as above. None reacted, either with the anti-MHC Class II (negative control) or with LAG-3 D1-D4 Ig, showing that the binding of LAG-3 D1-D4 is specific to MHC Class II molecules.

Further experiments were carried out using (i) mouse fibroblasts transfected or otherwise with genes coding for human DR7 or human DP4, (ii) mouse cells expressing or otherwise MHC Class II molecules, (iii) activated human CD4⁺ or CD8⁺ cells, and (iv) T lymphocyte lines expressing the different haplotypes of MHC Class II molecules (FIG. 8).

Unlike CD8 Ig, LAG-3 D1-D4 Ig binds to all cells expressing MHC Class II as efficiently as the anti-MHC Class II monoclonal antibody 949. LAG-3 D1-D4 Ig binds to all DR and DP haplotypes tested, to human MHC Class II molecules expressed by transfected mouse cells, to murine MHC Class II molecules and also to MHC Class II molecules expressed by CD4⁺ or CD8⁺ T lymphocytes.

These results represent for the first time proof that soluble molecules derived from a ligand for MHC Class II are capable of binding to cells expressing MHC Class II.

Similar experiments showed that LAG-3 D1D2 bound to cells expressing MHC Class II in as specific a manner and with the same efficiency as LAG-3 D1-D4.

Binding activity of LAG-3Ig and cellular distribution of ligands for LAG-3Ig

The capacity of this immunoadhesin to bind to cell ligands is measured using a fluorescein-labelled goat serum directed against human immunoglobulins.

In these experiments, the target cells are first incubated with a human monoclonal antibody or an immunoadhesin for 30 min at 4° C. in RPMI 1640 containing 10% of FCS (foetal calf serum). The cells are then incubated with an FITC-labelled goat anti-mouse immunoglobulin serum (Coulter) for the murine monoclonal antibodies or with an FITC-labelled goat anti-human immunoglobulin serum (Tago) for the immunoadhesins. The fluorescence is measured after two washes, analyzing 3,000 cells with an Elite cytometer (Coultronics, Hialeah, Fla.). FIG. 9 shows the degrees of binding of LAG-3Ig, CD8Ig, antibody 949 or antibody OKT3 (anti-CD3, ATCC), represented by the number of cells counted as a function of the logarithm of the measured fluorescence intensity.

LAG-3Ig binds to mouse fibroblasts transfected for the gene for the HLA DR₄ molecule, and does not bind to untransfected cells. CD8Ig is incapable of binding to HLA DR₄ ⁺ fibroblasts under the same conditions.

The cellular distribution of the ligands for LAG-3Ig was evaluated on a cell population sample by immunofluorescence.

LAG-3Ig is visualized on all positive Class II cells tested, including B cell lines transformed by Epstein-Barr virus (derived from genetically unrelated donors, including 10 homozygous lines of DR₁ to DR₁₀ typing), as well as on activated T and NK cells.

FIG. 9 shows, by way of example, the binding of LAG-3Ig to Daudi cells which are positive for Class II antigens.

The mean fluorescence intensity with LAG-3Ig is similar to that observed with antibody 949 which is specific for Class II antigens. The binding of LAG-3Ig to DR₄ (FIG. 9), DR₂, DR₇ or DPw4 (not shown) expressed at the surface of mouse fibroblasts is, in contrast, weaker than that observed for antibody 949.

No binding is detected on cell lines which are negative for Class II antigens of T origin (peripheral blood T cells, CEM, HSB2, REX lines), of B origin (RJ 2.2.5 line) or of non-lymphoid origin (human lines, K562 of erythromyoloid origin and line originating from melanoma cells (not shown)).

Moreover, LAG-3Ig binds to xenogeneic Class II molecules of the MHC, such as the antigens expressed by mouse lymphoid A 20 and the monkey Classes II expressed by phytohaemagglutinin-stimulated blasts (data not shown).

The specificity of binding of LAG-3Ig was also verified using the monoclonal antibodies 17B4, whose capacity to block LAG-3/MHC Class II interactions in cell adhesion tests was demonstrated beforehand (FIG. 10).

In these experiments, the LAG-3Ig molecules are preincubated for 30 minutes at 4° C. either with medium alone, or with 17B4 (1 mg/ml), or with OKT3 (1 mg/ml), before being brought into contact with Daudi cells.

FIG. 10 shows that a preincubation of LAG-3Ig with 17B4 inhibits the binding to Class II⁺ cells, whereas no inhibition is detected with the OKT3 control.

EXAMPLE 5 Inhibition of LAG-3/MHC Class II interaction by soluble fragments of LAG-3

The inhibition of LAG-3/MHC Class II interaction by the soluble fragments of LAG-3 may be observed directly in relation to the binding of LAG-3Ig by Class II MHC molecules, by competitive experiments with the soluble fragments.

To verify whether the soluble LAG-3D₁ D₂ fragments produced by CHO cells could displace the binding of immunoadhesins derived from LAG-3, the following tests were carried out:

Daudi cells are incubated with soluble LAG3-D₁ D₂ fragments so as to permit the binding of these molecules to the MHC Class II antigens expressed at the surface of the Daudi cells.

In a second step, the cells are incubated in the presence of LAG-3D₁ D₄ Ig in dimeric form or LAG-3D₁ D₂ Ig in monomeric form.

The binding of these immunoadhesins derived from LAG-3 is measured using a goat anti-human Ig F(ab')₂ conjugated to fluorescein (GAH-FITC).

The control groups are represented by Daudi cells incubated with dimeric LAG-3D₁ D₄ Ig or monomeric LAG-3D₁ D₂ Ig without preincubation with the soluble LAG-3D1D2 fragments.

The results are recorded in Table 5, which shows that the soluble LAG-3D₁ D₂ fragments are capable of displacing the immunoadhesins derived from LAG-3 in monoor dimeric form.

                  TABLE 5     ______________________________________                          Mean Fluor-     Reactants  Detection escence   Conclusion     ______________________________________     --         GAH-FITC  0.3       GAH does not                                    interfere     Dimeric    GAH-FITC  20.8      The binding of     LAG-3D1D4Ig                    CHO/LAG-3D1D2                                    inhibits the binding     CHO/LAG-3D1D2,                GAH-FITC  8.5       of dimeric     then dimeric                   LAG-3D1D4Ig (58%)     LAG-3D1D4Ig     Monomeric  GAH-FITC  62.5      The binding of     LAG-3D1D2Ig                    CHO/LAG-3D1D2                                    inhibits the binding     CHO/LAG-3D1D2,                GAH-FITC  10.9      of monomeric     then monomeric                 LAG-3D1D2Ig (27%)     LAG-3D1D2Ig     ______________________________________

These data confirm that the soluble LAG-3D1D2 fragments bind to MHC Class II molecules.

Inhibition of LAG-3/MHC Class II and CD4/MHC Class II interaction

Rosette formation between Cos cells transfected with wild-type LAG-3 and B lymphocytes transformed with EBV expressing MHC Class II molecules was demonstrated by BAIXERAS et al. (2). This interaction is inhibited both by anti-LAG-3 and anti-MHC Class II monoclonal antibodies.

The method described in this publication was modified by replacing the visualization and counting of Cos cells binding to B lymphocytes by counting the radioactivity remaining after incubation of ⁵¹ Cr-labelled B lymphocytes with Cos cells expressing LAG-3 (binding assay).

The possible inhibitory effects of soluble molecules derived from LAG-3 on LAG-3/MHC Class II interaction, and also on CD4/MHC Class II interaction, were studied.

Cos cells transfected with a suitable expression vector (coding for wild-type LAG-3 or for CD4). Two days later, the Cos cells were treated with trypsin and plated out again on the basis of 0.05×106 cells/well on flat-bottomed 12-well tissue culture plates. 24 hours later, ⁵¹ Cr-labelled Daudi cells (5.5×10⁶) were incubated on this monolayer of Cos cells (final vol.: 1 ml) for 1 hour. The target B cells were then aspirated off and the wells washed 5 to 7 times, gently adding 1 ml of medium dropwise. The edges of the wells were washed by suction using a Pasteur pipette. The remaining cells were lysed with 1 ml of PBS, 1% Triton for 15 minutes at 37° C. The lysates were centrifuged at 3000 rpm for 10 minutes, and 100 μl of the resulting supernatant were counted.

LAG-3 D1-D4 Ig was used to inhibit LAG-3/MHC Class II and CD4/MHC Class II interaction in the ⁵¹ Cr binding assay. Human CD8 Ig and IgG1 were tested in parallel and used as negative controls.

A significant inhibition of LAG-3/Class II interaction by LAG-3 D1-D4 Ig was detected (FIG. 5A). However, the LAG-3/MHC Class II interaction can be partially and non-specifically inhibited by human CD8 Ig and IgG1. Moreover, LAG-3 Ig proved to be a potential inhibitor of CD4/Class II interaction (FIG. 5B) under experimental conditions in which CD4/MHC Class II interaction was not modified by human CD8 Ig or IgG1. This suggests that LAG-3/Class II interaction is weaker than CD4/Class II interaction. These results represent the first proof of a possible competition of soluble molecules in an interaction of MHC Class II with its ligands.

EXAMPLE 6 Immunosuppressant activity of LAG-3 D1-D4 Ig

Functional tests were performed using the proliferation tests described above for the biological activity of the anti-LAG-3 monoclonal antibodies.

3 days and 5 days (D3 and D5) after antigenic stimulation, LAG-3 D1-D4 Ig showed a strong inhibition of the proliferation of clone 28, while human CD8 Ig and IgG had no effect (FIG. 6). Similar experiments were carried out with clone 154 (FIG. 7), and showed a partial inhibition in the presence of LAG-3 Ig. A control carried out with anti-LAG-3 monoclonal antibodies had the reverse effects, as observed previously.

A significant inhibition of the cell proliferation of cells incubated in the presence of LAG-3 D1-D4 Ig was also observed for clone 28.

These observations show that LAG-3 D1-D4 Ig is a potential immunosuppressant of the proliferation of T lymphocytes stimulated by an antigen, and indicate that LAG-3 might act as an "extinguisher" of the secondary immune response induced by activated CD4⁺ T helper lymphocytes.

Role of LAG-3Ig in the negative regulation of the immune responses of T cells

To demonstrate that a soluble form of LAG-3, mimicking the functions of the membrane molecule, could inhibit the activation of CD₄ ⁺ T clones stimulated by an antigen, the following tests were carried out on clone T154: the T cells are incubated beforehand with a saturating amount of LAG-3Ig (100 nM). The cells are then washed twice with cold RPMI and incubated with 10 μg/ml of goat antibodies directed against human immunoglobulins (Tago) at 4° C. for 30 minutes.

After two more washes, the cells are resuspended in RPMI containing 10% of foetal calf serum and incubated for 2 hours at 37° C. before adding the signal. To couple ("cross-link") the monoclonal antibodies, a goat anti-mouse antibody at a concentration of 10 μg/ml (Tago) is used.

FIG. 11 depicts an experiment in which clone T154 has been preincubated with LAG-3Ig bound ("cross-linked") to a second reactant (polyclonal serum specific for the constant region of human immunoglobulins). The degree of binding of LAG-3Ig to the cells is measured by immunofluorescence (FIG. 11A). FIG. 11B shows that a more than 50% inhibition of the proliferation of clone T154 is produced by LAG-3Ig. Under the same experimental conditions, no effect is observed with the control CD8Ig or with LAG-3Ig without "cross-linking" (not shown in the figure).

FIG. 11C also shows that no effect is observed when LAG-3Ig is used to bind ("cross-link") the MHC Class II molecules expressed by antigen-presenting B cells.

The possible effects of bound ("cross-linked") anti-Class II monoclonal antibodies in relation to the proliferation of T cells were compared to those of LAG-3Ig. A weak inhibition (less than 50%) is observed with antibody 949 and antibody D1.12 (anti-DR) bound to a goat anti-mouse polyclonal serum (FIG. 12). The inhibition of proliferation is hence epitope-dependent, the largest effect being obtained with the epitope of LAG-3 specific for the binding to Classes II.

The effects of LAG-3 Ig on the proliferation of T cells were also studied using different signals on another CD₄ ⁺ T clone, clone TDEL specific for peptide 34-53 of the basic myelin protein.

An inhibition of proliferation is observed (n=2) when TDEL is stimulated with the antigen (not shown), with immobilized OKT3 (FIG. 13A), with lectins (PHA+PMA) (FIG. 13B) and with 5 IU/ml of IL₂ (FIG. 13C). No inhibition is observed with 100 IU/ml of IL₂ (FIG. 13D).

In conclusion, these results collectively show that LAG-3 and MHC Class II molecules, which are each T cell-activating antigens, may be likened to effector molecules involved in the phase of inactivation of T cell responses. Moreover, these results illustrate the importance of interactions between T cells in the control of the cellular immune response.

EXAMPLE 7 Stimulation of cell cytotoxicity by LAG-3Ig

The role of LAG-3Ig in relation to cell cytotoxicity is studied on two types of effector cells:

freshly drawn human peripheral blood lymphocytes (PBL),

S1B5 line cells (clone of human NK cells).

The cytotoxic activity of these cells is measured by counting the ⁵¹ Cr released into the medium by previously labelled target cells, in the presence or absence of LAG-3Ig in the medium.

FIG. 14 shows the degree of cytotoxicity of S1B5 for a line of human B cells transformed by Epstein-Barr virus and carrying major histocompatibility complex Class I and II antigens (LAZ 388 line), as a function of different reactants added to the cultures.

Measurements are carried out after 4 hours of coculture for effector/target (S1B5/LAZ 388) cell ratios of 3:1 (clear columns) or 1:1 (shaded columns).

The negative controls consist of medium alone (MED), the immunoadhesin CD8Ig and the monoclonal antibody 17.B4 (anti-LAG-3).

The positive controls consist of three different monoclonal antibodies:

antibody L243 directed against Class II DR antigens,

antibody 9.49 directed against Class II DR, DP, DQ antigens,

antibody W632 directed against human major histocompatibility complex Class I antigens.

Anti-HLA Class I (W632) or Class II (L243) antibodies increase the lysis of the target cells (and not the 17B4 control). The immunoadhesin LAG-3Ig increases the lysis. The CD8Ig control has no effect.

FIG. 15 shows the results of an experiment similar to the above, in which the cytotoxicity of PBL with respect to Daudi cells (HLA Class I⁻) is measured, for effector/target ratios of 50:1 (clear columns) and 15:1 (shaded columns). The reactants added to the medium are the same as the ones used in the first experiment, except for antibody 9.49 and antibody 17.B4. Antibody 10H3 is an isotype IgG1 immunoglobulin specific for the CD45 surface antigen. It is used as negative control.

No change is observed with an antibody directed against major histocompatibility complex Class I antigens (W632).

The data from these two series of measurements show that, compared to negative controls, LAG-3Ig activates the cytotoxicity of NK cells. This effect is similar to the one observed with antibodies directed against MHC Class II molecules.

BIBLIOGRAPHIC REFERENCES

1. TRIEBEL T. et al., 1990, J. Exp. Med. 171, 1393-1405

2. BAIXERAS E. et al., 1992, J. Exp. Med. 176, 327-337

3. COSGROVE D. et al., 1991, Cell 66, 1051-1066

4. RAHEMTULLA A. et al., 1991, Nature 353, 180-184

5. TRAUNECKER A. et al., 1988, Nature 331, 84-86

6. BENEDICT A. A. et al., 1967, Methods in Immunology 1, 197-306 (1967)

7. YELTON D. E. et al., Ann. Rev. of Biochem. 50, 657-680 (1981)

8. HUARD B. et al., Immunogenetics 39: 213

9. MANIATIS T. et al. (1982), Molecular cloning: A laboratory manual--Cold Spring Harbor Laboratory, New-York.

10. SEED B., 1987, Nature 329, 840-842

11. COLE S. C. et al., Biotechnology 11, 1014-1024, 1993

12. COLE S. C. et al., Biotechnology 11, 1014-1024, 1993.

                                      TABLE NO. 1     __________________________________________________________________________                              reisdue                                    atom type,     Atom                     name and                                    charge and     name         x      y      z      no.   no.     __________________________________________________________________________     N   25.172370911                27.259836197                       67.855064392                              AP-n                                  40                                    n3                                      -0.5000                                           1     NN2 25.667764664                26.471963882                       67.420585632                              AP-n                                  40                                    hn                                       0.1300                                           2     CA  24.625223160                26.867494583                       69.180244446                              AP-n                                  40                                    ca                                       0.1200                                           3     NN1 24.393711090                27.474891663                       67.220008850                              AP-n                                  42                                    hn                                       0.1300                                           4     MA  23.936895370                27.680395126                       69.464080811                              AP-n                                  40                                    h  0.0700                                           5     C   25.662780726                26.773513794                       70.350120544                              AP-n                                  40                                    c'                                       0.3800                                           6     O   25.295415878                27.090747833                       71.482070923                              AP-n                                  40                                    o'                                      -0.4100                                           7     CB  23.766021729                25.587018967                       68.990669250                              AP-n                                  40                                    c2                                      -0.2600                                           8     MB1 23.060285568                25.72443695                       68.152351379                              AP-n                                  40                                    h  0.0700                                           9     MB2 24.413969040                24.744903564                       68.686981201                              AP-n                                  40                                    h  0.0700                                           10     CG  22.921775818                25.153419495                       70.196960449                              AP-n                                  40                                    c-                                       0.3400                                           11     DD1 22.069602966                25.929233551                       70.676017761                              AP-n                                  40                                    o-                                      -0.5700                                           12     DD2 23.115716934                24.009321213                       70.667663574                              AP-n                                  40                                    o-                                      -0.5700                                           13     M   26.906179428                26.304588318                       70.124969482                              SER 41                                    n -0.5000                                           14     CA  27.860145569                25.912786484                       71.207519531                              SER 41                                    ca                                       0.1200                                           15     HN  27.120641708                26.221813202                       69.126319885                              SER 41                                    hn                                       0.2800                                           16     MA  27.374551773                25.088045120                       71.766326904                              SER 41                                    h  0.1000                                           17     C   28.252065659                27.005065918                       72.271789551                              SER 41                                    c'                                       0.3800                                           18     O   27.987834930                28.200620651                       72.115295410                              SER 41                                    o'                                      -0.3800                                           19     CB  29.083601227                25.298025131                       70.494880676                              SER 41                                    c2                                      -0.1700                                           20     MB1 28.786190033                24.599395752                       69.690521240                              SER 41                                    h  0.1000                                           21     MB2 29.691858292                26.092912674                       70.004081726                              SER 41                                    h  0.1000                                           22     OG  29.905221939                24.578149796                       71.424118042                              SER 41                                    oh                                      -0.3800                                           23     HG  30.662555695                24.231645584                       70.939277649                              SER 41                                    ho                                       0.3500                                           24     N   28.857526779                26.558052063                       73.387054443                              GLY 42                                    n -0.5000                                           25     CA  29.199718475                27.440891266                       74.535232544                              GLY 42                                    cg                                       0.0200                                           26     MN  29.251720428                25.616510391                       73.267890930                              GLY 42                                    hn                                       0.2800                                           27     MA1 28.520187378                28.312047958                       74.591857910                              GLY 42                                    h  0.1000                                           28     MA2 28.983697891                26.890144348                       75.468444824                              GLY 42                                    h  0.1000                                           29     C   30.691051483                27.875793457                       74.601707458                              GLY 42                                    c'                                       0.3800                                           30     O   31.504199982                27.026502609                       74.980445862                              GLY 42                                    o'                                      -0.3800                                           31     N   31.113182068                29.132183075                       74.266487122                              PRO 43                                    n -0.4200                                           32     CA  32.858349609                29.476200104                       74.126792908                              PRO 43                                    ca                                       0.0600                                           33     HA  33.096603394                28.605407715                       73.708091736                              PRO 43                                    h  0.1000                                           34     CD  30.24075089                30.174203873                       73.722633362                              PRO 43                                    c2                                       0.0600                                           35     MD1 29.467987061                30.516298294                       74.466743469                              PRO 43                                    h  0.1000                                           36     MD2 29.664882660                29.799777985                       72.838768005                              PRO 43                                    h  0.1000                                           37     C   33.318023682                29.988374710                       75.414916992                              PRO 43                                    c'                                       0.3800                                           38     O   32.682361603                30.557033539                       76.312332153                              PRO 43                                    o'                                      -0.3800                                           39     CB  32.483139038                30.574260712                       73.043136597                              PRO 43                                    c2                                      -0.2000                                           40     MB1 33.350620270                31.263189316                       73.049049377                              PRO 43                                    h  0.1000                                           41     MB2 32.463790894                30.110534668                       72.036743164                              PRO 43                                    h  0.1000                                           42     CG  31.160949707                31.299722672                       73.307487488                              PRO 43                                    c2                                      -0.2000                                           43     MG1 31.279897690                32.024162292                       74.137779236                              PRO 43                                    h  0.1000                                           44     MG2 30.794561386                31.862808228                       72.428352356                              PRO 43                                    h  0.1000                                           45     K   34.683673859                29.902477264                       75.503486633                              PRO 44                                    n -0.4200                                           46     CA  35.485736847                30.679145813                       76.490524292                              PRO 44                                    ca                                       0.0600                                           47     KA  35.018527985                30.645456314                       77.491592407                              PRO 44                                    h  0.1000                                           48     CD  35.509281158                29.014368057                       74.655700684                              PRO 44                                    c2                                       0.0600                                           49     MD1 35.411357880                29.247959137                       73.577461243                              PRO 44                                    h  0.1000                                           50     MD2 35.214973450                27.955932617                       74.801994324                              PRO 44                                    h  0.1000                                           51     C   35.700843811                32.172924042                       76.063224792                              PRO 44                                    c'                                       0.3800                                           52     O   36.448230743                32.477428436                       75.126922607                              PRO 44                                    o'                                      -0.3800                                           53     CB  36.779544830                29.842718124                       76.547348022                              PRO 44                                    c2                                      -0.2000                                           54     HB1 37.662769315                30.430650711                       75.863670149                              PRO 44                                    h  0.1000                                           55     HB2 36.667564392                29.027103424                       77.288825989                              PRO 44                                    h  0.1000                                           56     CG  36.940769196                29.250995636                       75.143180867                              PRO 44                                    c2                                      -0.2000                                           57     MG1 37.446662903                29.982709885                       74.483322144                              PRO 44                                    h  0.1000                                           58     MG2 37.553295135                26.329860667                       75.134750366                              PRO 44                                    h  0.1000                                           59     N   35.026676178                33.104183197                       76.753837585                              ALA 45                                    h -0.5000                                           60     CA  35.034278870                34.544979095                       76.400695801                              ALA 45                                    ca                                       0.1200                                           61     MN  34.452354431                32.747509003                       77.536170959                              ALA 45                                    hn                                       0.2800                                           62     MA  35.105010986                34.659946442                       75.298950195                              ALA 45                                    h  0.1000                                           63     C   36.209384918                35.322113037                       77.076705833                              ALA 45                                    c'                                       0.3800                                           64     O   36.163528442                35.637268066                       78.268325806                              ALA 45                                    o'                                      -0.3800                                           65     CB  33.646369934                35.083190918                       76.800727844                              ALA 45                                    c3                                      -0.3000                                           66     MB1 33.534698486                36.150661469                       76.535202026                              ALA 45                                    h  0.1000                                           67     MB2 32.828392029                34.539138794                       76.290328979                              ALA 45                                    h  0.1000                                           68     MB3 33.465579987                35.001335144                       77.890525818                              ALA 45                                    h  0.1000                                           69     N   37.266757965                35.613758087                       76.297294617                              ALA 46                                    n -0.5000                                           70     MN  37.216701508                35.231555939                       75.346885681                              ALA 46                                    hn                                       0.2800                                           71     CA  38.489383698                36.310386658                       76.786270142                              ALA 46                                    ca                                       0.1200                                           72     MA  38.262126923                36.871456146                       77.716934204                              ALA 46                                    h  0.1000                                           73     C   39.058414459                37.311935425                       75.727844238                              ALA 46                                    c'                                       0.3800                                           74     O   38.922710419                37.100418091                       74.516731262                              ALA 46                                    o'                                      -0.3800                                           75     CB  39.526046753                35.215301514                       77.108406067                              ALA 46                                    c3                                      -0.3000                                           76     MB1 40.446556091                35.633434296                       77.555480957                              ALA 46                                    h  0.1000                                           77     MB2 39.131206512                34.469978333                       77.822120667                              ALA 46                                    h  0.1000                                           78     MB3 39.821903229                34.663463593                       76.197715759                              ALA 46                                    h  0.1000                                           79     N   39.737388611                38.384365082                       76.180320740                              ALA 47                                    n -0.5000                                           80     CA  40.295833588                39.429889679                       75.275863647                              ALA 47                                    ca                                       0.1200                                           81     MN  39.717037201                38.512592316                       77.196357727                              ALA 47                                    hn                                       0.2800                                           82     MA  39.901103973                39.319335938                       74.245994568                              ALA 47                                    h  0.1000                                           83     C   41.869789124                39.413467407                       75.170806885                              ALA 47                                    c'                                       0.3800                                           84     O   42.518333435                40.166862488                       75.906578044                              ALA 47                                    o'                                      -0.3800                                           85     CB  39.722030640                40.769996643                       75.786285400                              ALA 47                                    c3                                      -0.3000                                           86     MB1 40.045078278                41.611721039                       75.145561218                              ALA 47                                    h  0.1000                                           87     MB2 38.615306854                40.778987885                       75.787467957                              ALA 47                                    h  0.1000                                           88     MB3 40.059757233                41.007873535                       76.813346863                              ALA 47                                    h  0.1000                                           89     N   42.537422180                38.621597290                       74.274406433                              PRO 48                                    n -0.4200                                           90     CA  44.009857178                38.718185425                       74.045745850                              PRO 48                                    ca                                       0.0600                                           91     MA  44.539390564                38.855972290                       75.008773804                              PRO 48                                    h  0.1000                                           92     CD  41.903011322                37.522361755                       73.516281128                              PRO 48                                    c2                                       0.0600                                           93     MD1 41.081176758                37.871139526                       72.860626221                              PRO 48                                    h  0.1000                                           94     MD2 41.490711312                36.761222839                       74.206848145                              PRO 48                                    h  0.1000                                           95     C   44.448683322                39.844142914                       73.044319153                              PRO 48                                    c'                                       0.3800                                           96     O   43.693031311                40.225761414                       72.144134521                              PRO 48                                    o'                                      -0.3800                                           97     CB  44.302902771                37.304885864                       73.500595093                              PRO 48                                    c2                                      -0.2000                                           98     MB1 45.227554321                37.245840894                       72.897834778                              PRO 48                                    h  0.1000                                           99     MB2 44.442562103                36.597515106                       74.341529846                              PRO 48                                    h  0.1000                                          100     CG  43.051033020                36.925685883                       72.702011108                              PRO 48                                    c2                                      -0.2000                                          101     MG1 43.084365845                37.389282227                       71.696128845                              PRO 48                                    h  0.1000                                          102     MG2 42.948661804                35.835418701                       72.555740356                              PRO 48                                    h  0.1000                                          103     N   45.700798035                40.324272156                       73.165817261                              GLY 49                                    n -0.5000                                          104     CA  46.289730072                41.291931152                       72.184875488                              GLY 49                                    cg                                       0.0200                                          105     MN  46.207077026                39.986907959                       73.991729736                              GLY 49                                    hn                                       0.2800                                          106     MA1 45.620616913                41.460151672                       71.317451477                              GLY 49                                    h  0.1000                                          107     MA2 46.357063293                42.283206940                       72.672386169                              GLY 49                                    h  0.1000                                          108     C   47.682319641                40.950855255                       71.600997925                              GLY 49                                    c'                                       0.3800                                          109     O   48.560806274                41.811416626                       71.601501465                              GLY 49                                    o'                                      -0.3800                                          110     N   47.842975616                39.718383789                       71.091018677                              HIS 50                                    n -0.5000                                          111     MN  46.947166443                39.222202301                       71.052452087                              HIS 50                                    hn                                       0.2800                                          112     CA  49.020114899                39.216835022                       70.306198120                              HIS 50                                    ca                                       0.1200                                          113     MA  49.324539185                38.296161652                       70.836235046                              HIS 50                                    h  0.1000                                          114     C   50.400863647                39.993576050                       70.182189941                              HIS 50                                    c'                                       0.3800                                          115     O   50.733478546                40.446022034                       69.081466675                              HIS 50                                    o'                                      -0.3800                                          116     CB  48.455345154                38.673969269                       68.953773499                              HIS 50                                    c2                                      -0.2000                                          117     HB1 49.238502502                38.051708221                       68.481765747                              HIS 50                                    h  0.1000                                          118     HB2 47.639427185                37.951225281                       69.144165039                              HIS 50                                    h  0.1000                                          119     CG  47.954322815                39.695293427                       67.919425964                              HIS 50                                    c5                                       0.1000                                          120     MD1 46.759342194                40.404747009                       68.035293579                              HIS 50                                    np                                      -0.4200                                          121     CE1 46.825572968                41.133701324                       66.873245239                              HIS 50                                    c5                                       0.2700                                          122     NE2 47.887611389                40.964004517                       66.014495850                              HIS 50                                    np                                      -0.5000                                          123     CD2 48.617565155                40.019775391                       66.717041016                              HIS 50                                    c5                                       0.0100                                          124     ME1 46.043247223                41.852840424                       66.655242920                              HIS 50                                    h  0.1300                                          125     ME2 48.092224121                41.403381348                       65.108955383                              HIS 50                                    hn                                       0.2800                                          126     MD2 49.566501617                39.599807739                       66.396347046                              HIS 50                                    h  0.1300                                          127     N   51.278770447                40.105480194                       71.226615906                              PRO 51                                    n -0.4200                                          128     CA  52.667034149                40.618915558                       71.077911377                              PRO 51                                    ca                                       0.0600                                          129     MA  52.723186493                41.393623352                       70.287094116                              PRO 51                                    h  0.1000                                          130     CD  50.956447601                39.767082214                       72.624496460                              PRO 51                                    c2                                       0.0600                                          131     MD1 50.920970917                38.670558929                       72.747383118                              PRO 51                                    h  0.1000                                          132     MD2 49.988422394                40.190521240                       72.947166443                              PRO 51                                    h  0.1000                                          133     C   53.707103729                39.478122711                       70.804489136                              PRO 51                                    c'                                       0.3800                                          134     O   53.623699188                38.391666412                       71.391418457                              PRO 51                                    o'                                      -0.3800                                          135     CB  52.846694946                41.283794403                       72.455955505                              PRO 51                                    c2                                      -0.2000                                          136     MB1 53.907955170                41.442169189                       72.729286194                              PRO 51                                    h  0.1000                                          137     MB2 52.373264313                42.285846710                       72.449127197                              PRO 51                                    h  0.1000                                          138     CG  52.105823517                40.370025635                       73.440398170                              PRO 51                                    c2                                      -0.2000                                          139     MG1 52.782051086                39.565395355                       73.789718628                              PRO 51                                    h  0.1000                                          140     MG2 51.753723145                40.912036896                       74.337333679                              PRO 51                                    h  0.1000                                          141     N   54.704883575                39.729522705                       69.933471680                              LEU 52                                    n -0.5000                                          142     CA  55.791530609                38.746330261                       69.603820801                              LEU 52                                    ca                                       0.1200                                          143     MN  54.653743744                40.652229308                       69.490356445                              LEU 52                                    hn                                       0.2800                                          144     MA  56.479202271                39.288208008                       68.927917480                              LEU 52                                    h  0.1000                                          145     C   35.301837921                37.525024414                       68.745040894                              LEU 52                                    c'                                       0.3800                                          146     O   55.637695313                37.425930023                       67.562049866                              LEU 52                                    o'                                      -0.3800                                          147     CB  56.671585083                38.316761017                       70.829437256                              LEU 52                                    c2                                      -0.2000                                          148     MB1 56.036743164                37.710464478                       71.502593994                              LEU 52                                    h  0.1000                                          149     MB2 57.445541382                37.602729797                       70.487434387                              LEU 52                                    h  0.1000                                          150     CG  57.363307953                39.420749664                       71.675102234                              LEU 52                                    c1                                      -0.1000                                          151     MG  56.617557526                40.200679779                       71.926834106                              LEU 52                                    h  0.1000                                          152     CD1 57.875057220                38.833915710                       73.004547119                              LEU 52                                    c3                                      -0.3000                                          153     MD11         58.353130341                39.601875305                       73.642135620                              LEU 52                                    h  0.1000                                          154     MD12         57.048751831                38.403957367                       73.601577759                              LEU 52                                    h  0.1000                                          155     MD13         58.618564606                38.028480530                       72.852462769                              LEU 52                                    h  0.1000                                          156     CD2 58.531608582                40.085853577                       70.927200317                              LEU 52                                    c3                                      -0.3000                                          157     MD21         59.028976440                40.857166290                       71.545303345                              LEU 52                                    h  0.1000                                          158     MD22         59.309509277                39.355545044                       70.634819031                              LEU 52                                    h  0.1000                                          159     MD23         58.192760468                40.592193604                       70.005569458                              LEU 52                                    h  0.1000                                          160     N   54.534648895                36.601863861                       69.354270935                              ALA 53                                    h -0.5000                                          161     CA  53.940563202                35.420303345                       68.673507690                              ALA 53                                    ca                                       0.1200                                          162     MN  54.159572601                36.958141327                       70.246543884                              ALA 53                                    hn                                       0.2800                                          163     HA  53.600482941                35.753322601                       67.671340942                              ALA 53                                    h  0.1000                                          164     C   52.639778137                34.883575439                       69.383995056                              ALA 53                                    c'                                       0.3800                                          165     O   51.628326416                34.818698883                       68.677047729                              ALA 53                                    o'                                      -0.3800                                          166     CB  55.008785248                34.330768585                       68.423698425                              ALA 53                                    c3                                      -0.3000                                          167     MB1 54.582756042                33.460170746                       67.892036438                              ALA 53                                    h  0.1000                                          168     MB2 55.828662872                34.711517334                       67.787284851                              ALA 53                                    h  0.1000                                          169     MB3 55.479701896                33.961406708                       69.351325989                              ALA 53                                    h  0.1000                                          170     N   52.555103302                34.484893799                       70.698265076                              PRO 54                                    n -0.4200                                          171     CA  51.304950714                33.917499542                       71.286506653                              PRO 54                                    ca                                       0.0600                                          172     MA  50.878768921                33.172523499                       70.584587097                              PRO 54                                    h  0.1000                                          173     CD  53.705833435                34.435420990                       71.626182556                              PRO 54                                    c2                                       0.0600                                          174     MD1 54.215991974                35.409980774                       71.739936829                              PRO 54                                    h  0.1000                                          175     MD2 54.453365326                33.693523407                       71.288909912                              PRO 54                                    h  0.1000                                          176     C   50.177021027                34.945980072                       71.642837524                              PRO 54                                    c'                                       0.3800                                          177     O   50.377983093                36.164409637                       71.677795410                              PRO 54                                    o'                                      -0.3800                                          178     CB  51.867893219                33.173828125                       72.519187927                              PRO 54                                    c2                                      -0.2000                                          179     MB1 51.135505676                33.051830292                       73.340660095                              PRO 54                                    h  0.1000                                          180     MB2 52.181377411                32.150829315                       72.232276917                              PRO 54                                    h  0.1000                                          181     CG  53.087123871                33.989192963                       72.949317932                              PRO 54                                    c2                                      -0.2000                                          182     MG1 52.768703461                34.871643066                       73.538475037                              PRO 54                                    h  0.1000                                          183     MG2 53.791828156                33.414070129                       73.579086304                              PRO 54                                    h  0.1000                                          184     N   48.977436066                34.414421082                       71.936706543                              GLY 55                                    n  0.5000                                          185     CA  47.822620392                35.225021362                       72.404747009                              GLY 55                                    cg                                       0.0200                                          186     MN  48.958133698                33.389709473                       71.929512024                              GLY 55                                    hn                                       0.2800                                          187     MA1 47.830284119                36.238574982                       71.983676453                              GLY 55                                    h  0.1000                                          188     MA2 46.896526337                34.772380829                       72.004432678                              GLY 55                                    h  0.1000                                          189     C   47.670829773                35.263648987                       73.950416565                              GLY 55                                    c'                                       0.3800                                          190     O   47.247509003                34.242198944                       74.498336792                              GLY 55                                    o'                                      -0.3800                                          191     N   47.956153870                36.372848511                       74.696281433                              PRO 56                                    n -0.4200                                          192     CA  47.830066681                36.396587372                       76.179130554                              PRO 56                                    ca                                       0.0600                                          193     MA  48.225147247                35.457695007                       76.619560242                              PRO 56                                    h  0.1000                                          194     CD  48.653686523                37.556163788                       74.163940430                              PRO 56                                    c2                                       0.0600                                          195     MD1 48.108860016                38.040233612                       73.332565308                              PRO 56                                    h  0.1000                                          196     MD2 49.652721405                37.256084442                       73.799308777                              PRO 56                                    h  0.1000                                          197     C   46.361907959                36.804877472                       76.668052673                              PRO 56                                    c'                                       0.3800                                          198     O   45.890090942                37.732730865                       76.845039368                              PRO 56                                    o'                                      -0.3800                                          199     CB  48.804897308                37.531764984                       76.560951233                              PRO 56                                    c2                                      -0.2000                                          200     MB1 48.542221069                38.034294128                       77.511970520                              PRO 56                                    h  0.1000                                          201     MB2 49.825592041                37.124942236                       76.697402954                              PRO 56                                    h  0.1000                                          202     CG  48.782566071                38.488700867                       75.368530273                              PRO 56                                    c2                                      -0.2000                                          203     MG1 47.903289795                39.158111572                       75.434867859                              PRO 56                                    h  0.1000                                          204     MG2 49.679012299                39.133480072                       75.321792603                              PRO 56                                    h  0.1000                                          205     N   45.650573730                35.488880157                       76.896896362                              HIS 57                                    n -0.5000                                          206     MN  46.046119690                34.657379150                       76.439048767                              HIS 57                                    hn                                       0.2800                                          207     CA  44.244419098                35.504474640                       77.387596130                              HIS 57                                    ca                                       0.1200                                          208     MA  43.667560577                36.207649231                       76.759368896                              HIS 57                                    h  0.1000                                          209     C   44.173530579                35.943927765                       78.901000977                              HIS 57                                    c'                                       0.3800                                          210     O   44.750030518                35.234142303                       79.736030579                              HIS 57                                    o'                                      -0.3800                                          211     CB  43.610416412                34.095542908                       77.185058594                              HIS 57                                    c2                                      -0.2000                                          212     MB1 44.270858765                33.323795319                       77.623748779                              HIS 57                                    h  0.1000                                          213     MB2 42.689029694                34.034877777                       77.796188354                              HIS 57                                    h  0.1000                                          214     CB  43.250400543                33.671833038                       75.751731873                              HIS 57                                    c5                                       0.1000                                          215     MD1 44.116428375                33.723646257                       74.666069031                              HIS 57                                    np                                      -0.4200                                          216     CE1 43.325973511                33.139041901                       73.713264465                              HIS 57                                    c5                                       0.2700                                          217     ME2 42.066158295                32.716590881                       74.036811829                              HIS 57                                    np                                      -0.5000                                          218     CD2 42.045005798                33.048488617                       75.379264832                              HIS 57                                    c5                                       0.0100                                          219     ME1 43.722070465                33.001682281                       72.711807251                              HIS 57                                    h  0.1300                                          220     ME2 41.370864868                32.229383759                       73.464263916                              HIS 57                                    hn                                       0.3800                                          221     MD2 41.236827850                32.815635681                       76.058380127                              HIS 57                                    h  0.1300                                          222     N   43.513858795                37.068405151                       79.318038940                              PRO 58                                    n -0.4200                                          223     CA  43.593978882                37.569736481                       80.719970703                              PRO 58                                    ca                                       0.0600                                          224     MA  44.657642365                37.630668640                       81.026939392                              PRO 58                                    h  0.1000                                          225     CD  42.833599091                38.010932922                       78.407653809                              PRO 58                                    c2                                       0.0600                                          226     MD1 42.093353271                37.516807556                       77.751007080                              PRO 58                                    h  0.1000                                          227     MD2 43.569110870                38.527889252                       77.758674622                              PRO 58                                    h  0.1000                                          228     C   42.805988312                36.695770264                       81.747009277                              PRO 58                                    c'                                       0.3800                                          229     O   41.569808960                36.693984985                       81.769706726                              PRO 58                                    o'                                      -0.3800                                          230     CB  43.072513580                39.016571045                       80.579101563                              PRO 58                                    c2                                      -0.2000                                          231     MB1 42.559799194                39.392253876                       81.485702515                              PRO 58                                    h  0.1000                                          232     MB2 43.920654297                39.706352234                       80.398040771                              PRO 58                                    h  0.1000                                          233     CG  42.156440735                38.999496460                       79.353309631                              PRO 58                                    c2                                      -0.2000                                          234     MG1 41.151851654                38.629653931                       79.634307861                              PRO 58                                    h  0.1000                                          235     MG2 42.023620605                39.998752594                       78.896965027                              PRO 58                                    h  0.1000                                          236     N   43.540618896                35.961261749                       82.605430603                              ALA 59                                    n -0.5000                                          237     MN  44.537330627                35.932937622                       82.365699768                              ALA 59                                    hn                                       0.2800                                          238     CA  42.949653625                34.946208954                       83.526855469                              ALA 59                                    ca                                       0.1200                                          239     MA  42.197692871                34.355514526                       82.965911865                              ALA 59                                    h  0.1000                                          240     C   42.208984375                35.488883972                       84.803985596                              ALA 59                                    c'                                       0.3800                                          241     O   42.496433258                35.127353668                       85.948333740                              ALA 59                                    o'                                      -0.3800                                          242     CB  44.107444763                33.975612640                       83.839767456                              ALA 59                                    c3                                      -0.3000                                          243     MB1 43.761249542                33.130538940                       84.463287354                              ALA 59                                    h  0.1000                                          244     MB2 44.544620514                33.531585693                       82.924507141                              ALA 59                                    h  0.1000                                          245     MB3 44.925910950                34.467308044                       84.399597168                              ALA 59                                    h  0.1000                                          246     N   41.192062378                36.323741913                       84.559051514                              ALA 60                                    n -0.5000                                          247     MN  41.124549866                36.554992676                       83.555343628                              ALA 60                                    hn                                       0.2800                                          248     CA  40.203086853                36.790748596                       85.562988281                              ALA 60                                    ca                                       0.1200                                          249     MA  40.023948669                35.965785980                       86.283721924                              ALA 60                                    h  0.1000                                          250     C   38.823528290                37.027305602                       84.846977234                              ALA 60                                    c'                                       0.3800                                          251     O   37.897804260                36.283885956                       85.186340332                              ALA 60                                    o'                                      -0.3800                                          252     CB  40.756233215                37.971691132                       86.389617920                              ALA 60                                    c3                                      -0.3000                                          253     MB1 40.004146578                38.357006073                       87.102043152                              ALA 60                                    h  0.1000                                          254     MB2 41.631908417                37.656120300                       86.987319946                              ALA 60                                    h  0.1000                                          255     MB3 41.090072632                38.818138123                       85.765472412                              ALA 60                                    h  0.1000                                          256     N   38.616340637                37.921970367                       83.823921204                              PRO 61                                    n -0.4200                                          257     CA  37.403762817                37.874496460                       82.948486328                              PRO 61                                    ca                                       0.0600                                          258     MA  36.491020203                37.824863434                       83.573593140                              PRO 61                                    h  0.1000                                          259     CD  39.556003571                39.003845215                       83.459777832                              PRO 61                                    c2                                       0.0600                                          260     MD1 40.594814301                38.658184052                       83.304046631                              PRO 61                                    h  0.1000                                          261     MD2 39.571029663                39.778694153                       84.250648498                              PRO 61                                    h  0.1000                                          262     C   37.275394440                36.712963104                       81.892074585                              PRO 61                                    c'                                       0.3800                                          263     O   36.366387939                36.670307159                       81.183484568                              PRO 61                                    o'                                      -0.3800                                          264     CB  37.473857880                39.266963959                       82.286987305                              PRO 61                                    c2                                      -0.2000                                          265     MB1 36.949714661                39.321308136                       81.312759399                              PRO 61                                    h  0.1000                                          266     MB2 36.985511780                40.017913818                       82.938987732                              PRO 61                                    h  0.1000                                          267     CG  38.968631744                39.571613312                       82.168632507                              PRO 61                                    c2                                      -0.2000                                          268     HG1 39.397396088                39.046298981                       81.292793274                              PRO 61                                    h  0.1000                                          269     HG2 39.180202484                40.649600983                       82.039207458                              PRO 61                                    h  0.1000                                          270     N   38.242324829                35.777751923                       81.785034180                              SER 62                                    n -0.5000                                          271     CA  38.186004639                34.624855042                       80.844123840                              SER 62                                    ca                                       0.1200                                          272     MN  39.035541534                35.827360535                       82.416992188                              SER 62                                    hn                                       0.2800                                          273     MA  37.921787262                35.006183624                       79.841125488                              SER 62                                    h  0.1000                                          274     C   37.145660400                33.532897949                       81.261909485                              SER 62                                    c'                                       0.3800                                          275     O   37.466091156                32.626064301                       82.041137695                              SER 62                                    o'                                      -0.3800                                          276     CB  39.620265961                34.048240662                       80.725196838                              SER 62                                    c2                                      -0.1700                                          277     MB1 39.660293579                33.306644440                       79.904281616                              SER 62                                    h  0.1000                                          278     MB2 40.349685669                34.832687378                       80.442802429                              SER 62                                    h  0.1000                                          279     OG  40.032703400                33.414188385                       81.938880920                              SER 62                                    oh                                      -0.3800                                          280     MG  39.252223969                32.931293488                       82.256263733                              SER 62                                    ho                                       0.3500                                          281     N   35.902244568                33.647918701                       80.764541626                              SER 63                                    n -0.5000                                          282     CA  34.747528076                32.87465942                       81.297317505                              SER 63                                    ca                                       0.1200                                          283     MN  35.768447876                34.503852844                       80.205200195                              SER 63                                    hn                                       0.2800                                          284     MA  35.064254761                32.265518188                       82.170570374                              SER 63                                    h  0.1000                                          285     C   34.106758118                31.936998367                       80.231674194                              SER 63                                    c'                                       0.3800                                          286     O   33.716896057                32.367130280                       79.142120361                              SER 63                                    o'                                      -0.3800                                          287     CB  33.716815948                33.889484406                       81.843544006                              SER 63                                    c2                                      -0.1700                                          288     MB1 34.199871063                34.571384430                       82.572105408                              SER 63                                    h  0.1000                                          289     MB2 33.32880719                34.543502808                       81.036796570                              SER 63                                    h  0.1000                                          290     OG  32.634590149                33.222091675                       82.496467590                              SER 63                                    oh                                      -0.3800                                          291     MG  32.159793854                32.710407257                       81.832328796                              SER 63                                    ho                                       0.3500                                          292     N   33.914913177                30.658897400                       80.588378906                              TRP 64                                    n -0.5000                                          293     CA  33.112319946                29.694124222                       79.783546448                              TRP 64                                    ca                                       0.1200                                          294     MN  34.221500397                30.422899246                       81.538955688                              TRP 64                                    hn                                       0.2800                                          295     MA  33.404731750                29.812168121                       78.721931458                              TRP 64                                    h  0.1000                                          296     C   31.573041916                29.961977005                       79.883049011                              TRP 64                                    c'                                       0.3800                                          297     O   30.996980667                29.940891266                       80.975959778                              TRP 64                                    o'                                      -0.3800                                          298     CB  33.525466919                26.235471725                       80.142707825                              TRP 64                                    c2                                      -0.2000                                          299     MB1 32.950366974                27.534402847                       79.508041382                              TRP 64                                    h  0.1000                                          300     MB2 34.571674347                28.078489304                       79.816658020                              TRP 64                                    h  0.1000                                          301     CG  33.405326843                27.783784866                       81.611763000                              TRP 64                                    c5                                       0.0000                                          302     CD1 32.267101288                27.214570999                       82.221214294                              TRP 64                                    c5                                       0.1000                                          303     NE1 32.481933594                26.953943253                       83.590408325                              TRP 64                                    np                                      -0.5000                                          304     CE2 33.781982422                27.378627777                       83.812339783                              TRP 64                                    c5                                       0.1100                                          305     CD2 34.355152130                27.881061554                       82.617965698                              TRP 64                                    c5                                       0.0000                                          306     MD1 31.322025528                27.036033630                       81.708480835                              TRP 64                                    h  0.1000                                          307     ME1 31.820940018                26.578481674                       84.279945374                              TRP 64                                    hn                                       0.2800                                          308     CE3 35.681034088                28.394184113                       82.615905762                              TRP 64                                    cp                                      -0.1000                                          309     ME3 36.128986359                28.784986496                       81.714393616                              TRP 64                                    h  0.1000                                          310     CE3 36.396430969                28.387191772                       83.815704346                              TRP 64                                    cp                                      -0.1000                                          311     ME3 37.405311584                28.776082993                       83.832160950                              TRP 64                                    h  0.1000                                          312     CM2 35.830604553                27.888952255                       84.994010925                              TRP 64                                    cp                                      -0.1000                                          313     MM2 36.410472870                27.896844844                       85.906951904                              TRP 64                                    h  0.1000                                          314     CZ2 34.527233124                27.382776260                       85.014289856                              TRP 64                                    cp                                      -0.1000                                          315     MZ2 34.097515106                27.006088257                       85.929389954                              TRP 64                                    h  0.1000                                          316     N   30.921349498                30.232547760                       78.740600586                              GLY 65                                    n -0.5000                                          317     CA  29.460748672                30.504768372                       78.692192078                              GLY 65                                    cg                                       0.0200                                          318     MN  31.520374298                30.302478790                       77.901519775                              GLY 65                                    hn                                       0.2800                                          319     MA1 29.073087692                30.896234512                       79.650825500                              GLY 65                                    h  0.1000                                          320     MA2 29.288171768                31.333106995                       77.981094360                              GLY 65                                    h  0.1000                                          321     C   28.633579254                29.293350220                       78.197364807                              GLY 65                                    c'                                       0.3800                                          322     O   28.866907883                29.137302399                       76.975486755                              GLY 65                                    o'                                      -0.3800                                          323     N   27.969013672                28.429246902                       79.038352966                              PRO 66                                    n -0.4200                                          324     CA  27.282257080                27.212890625                       78.846752930                              PRO 66                                    ca                                       0.0600                                          325     MA  27.989650726                26.634012222                       77.917152405                              PRO 66                                    h  0.1000                                          326     CD  28.016592026                28.832529831                       80.511337280                              PRO 66                                    c2                                       0.0600                                          327     MD1 27.731332779                29.836006927                       80.880874634                              PRO 66                                    h  0.1000                                          328     MD2 29.027824402                28.301166534                       80.897590637                              PRO 66                                    h  0.1000                                          329     C   25.977466583                27.479314804                       77.725341797                              PRO 66                                    c'                                       0.3800                                          330     O   25.217950821                28.417282104                       77.982574463                              PRO 66                                    o'                                      -0.3800                                          331     CB  27.045602798                26.422634125                       79.851196289                              PRO 66                                    c2                                      -0.2000                                          332     MB1 26.132562637                25.797079086                       79.827728271                              PRO 66                                    h  0.1000                                          333     MB2 27.890687943                25.729280472                       80.029365540                              PRO 66                                    h -0.1000                                          334     CG  27.003501892                27.477243423                       80.958015442                              PRO 66                                    c2                                      -0.2000                                          335     MG1 25.990892410                27.921483994                       81.014678955                              PRO 66                                    h  0.1000                                          336     MH2 27.232566833                27.061700821                       81.956459045                              PRO 66                                    h  0.1000                                          337     N   25.734319687                26.626403809                       76.719802856                              ARG+                                  67                                    n -0.5000                                          338     CA  24.603988647                26.793025970                       75.767257690                              ARG+                                  67                                    ca                                       0.1200                                          339     MN  26.386735916                25.841371536                       76.649803162                              ARG+                                  67                                    hn                                       0.2800                                          340     MA  24.496238708                27.874828339                       75.561668396                              ARG+                                  67                                    h  0.1000                                          341     C   23.227464676                26.224872589                       76.267372131                              ARG+                                  67                                    c'                                       0.2800                                          342     O   23.178310394                25.034952164                       76.603759766                              ARG+                                  67                                    o'                                      -0.3800                                          343     CB  24.990058899                26.165229797                       74.398826599                              ARG+                                  67                                    c2                                      -0.2000                                          344     MB1 24.135663986                26.318639755                       73.709175110                              ARG+                                  67                                    h  0.1100                                          345     MB2 25.787433624                26.779323578                       73.940032959                              ARG+                                  67                                    h  0.1100                                          346     CG  25.439929962                24.676465988                       74.361564636                              ARG+                                  67                                    c2                                      -0.2000                                          347     MG1 26.546255112                24.646316528                       74.415458679                              ARG+                                  67                                    h  0.1300                                          348     MG2 25.092346191                24.131168365                       75.261718750                              ARG+                                  67                                    h  0.1300                                          349     CD  24.934387207                23.941221237                       73.112297058                              ARG+                                  67                                    c2                                      -0.0900                                          350     MD  23.838283539                23.774566650                       73.188652039                              ARG+                                  67                                    h  0.1300                                          351     MD2 25.070211411                24.585262299                       72.220893860                              ARG+                                  67                                    h  0.1300                                          352     ME  25.665744781                22.657058716                       72.968780518                              ARG+                                  67                                    n1                                      -0.5000                                          353     ME  26.251846313                22.313375473                       73.731925964                              ARG+                                  67                                    hn                                       0.3600                                          354     CZ  25.689014435                21.902687073                       71.871635437                              ARG+                                  67                                    cr                                       0.4500                                          355     MM1 26.493299484                20.879484177                       71.859733582                              ARG+                                  67                                    n2                                      -0.5000                                          356     M11 27.072875705                20.740955353                       72.690315247                              ARG+                                  67                                    hn                                       0.3600                                          357     MM12         26.520929337                20.119580078                       71.006805420                              ARG+                                  67                                    hn                                       0.3600                                          358     MM2 24.956668854                22.117029190                       70.805320740                              ARG+                                  67                                    n2                                      -0.5000                                          359     MM21         25.030595779                21.456792831                       70.033142090                              ARG+                                  67                                    hn                                       0.3600                                          360     MM22         24.266489029                22.916227341                       70.850784302                              ARG+                                  67                                    hn                                       0.3600                                          361     N   22.080270767                26.971176147                       76.244255066                              ARG+                                  68                                    n -0.4200                                          362     CA  20.743734360                24.358839035                       76.485237122                              PRO 68                                    ca                                       0.0600                                          363     MA  20.817556381                25.605155945                       77.294357300                              PRO 68                                    h  0.1000                                          364     CD  22.076143265                28.448001862                       76.342918396                              PRO 68                                    c2                                       0.0600                                          365     MD1 22.539228439                28.949869156                       75.469612122                              PRO 68                                    h  0.1000                                          366     MD2 22.632146835                28.776126862                       77.244499207                              PRO 68                                    h  0.1000                                          367     C   20.182382584                25.586324692                       75.240180969                              PRO 68                                    c'                                       0.3800                                          368     O   20.420539856                24.381649017                       75.139877319                              PRO 68                                    o'                                      -0.3800                                          369     CB  19.948141098                27.549791336                       77.062515259                              PRO 68                                    c2                                      -0.2000                                          370     MB1 18.858430862                27.490163803                       76.877128601                              PRO 68                                    h  0.1000                                          371     MB2 20.066789627                27.567596436                       78.163002014                              PRO 68                                    h  0.1000                                          372     CG  20.592071533                28.804227829                       76.667758179                              PRO 68                                    c2                                      -0.2000                                          373     MG1 20.158363342                29.031393051                       75.478172302                              PRO 68                                    h  0.1000                                          374     MG2 20.428398132                29.704229355                       77.092338562                              PRO 68                                    h  0.1000                                          375     N   19.458055496                26.229068756                       74.300010681                              ARG+                                  69                                    n -0.5000                                          376     CA  18.893756866                25.542047501                       73.096710205                              ARG+                                  69                                    ca                                       0.1200                                          377     MN  19.221296310                27.198928833                       74.529014587                              ARG+                                  69                                    hn                                       0.2800                                          378     MA  19.667610168                24.872461319                       72.660797119                              ARG+                                  69                                    h  0.1000                                          379     C   18.514461517                26.608341217                       72.012504578                              ARG+                                  69                                    c'                                       0.3800                                          380     O   17.383218765                27.099323273                       72.036094666                              ARG+                                  69                                    o'                                      -0.3800                                          381     CB  17.681777954                24.654710770                       73.539657593                              ARG+                                  69                                    c2                                      -0.2000                                          382     MB1 18.011884689                23.935596466                       74.315147400                              ARG+                                  69                                    h  0.1100                                          383     MB2 16.954420090                25.310214996                       74.061965942                              ARG+                                  69                                    h  0.100                                          384     CG  16.946891785                23.862810135                       72.427490234                              ARG+                                  69                                    c2                                      -0.2000                                          385     MG1 16.649517059                24.851628113                       71.612152100                              ARG+                                  69                                    h  0.1300                                          386     MG2 17.638776779                23.127803802                       71.968805054                              ARG+                                  69                                    h  0.1300                                          387     CD  15.688191414                23.166488647                       72.975959778                              ARG+                                  69                                    c2                                      -0.0900                                          388     MD1 15.980383873                22.365550995                       73.686569214                              ARG+                                  69                                    h  0.1300                                          389     MD2 15.090404510                23.898859024                       73.554351807                              ARG+                                  69                                    h  0.1300                                          390     ME  14.889810562                22.612804413                       71.848815918                              ARG+                                  69                                    n1                                      -0.5000                                          391     ME  15.272388458                22.616128922                       70.898582458                              ARG+                                  69                                    hn                                       0.3600                                          392     CZ  13.644455910                22.143890381                       71.942466736                              ARG+                                  69                                    cr                                       0.4500                                          393     MM1 13.048615456                21.755460739                       70.850708008                              ARG+                                  69                                    n2                                      -0.5000                                          394     MM11         13.576630592                21.832328796                       69.979103088                              ARG+                                  69                                    hn                                       0.3600                                          395     MM12         12.090744019                21.411947250                       70.936080933                              ARG+                                  69                                    hn                                       0.3600                                          396     MM2 12.989639282                22.056533813                       73.074882507                              ARG+                                  69                                    n2                                      -0.5000                                          397     CZ  13.644455910                22.143890381                       71.942466736                              ARG+                                  69                                    cr                                       0.4500                                          393     MM1 13.048615456                21.755460739                       70.850708008                              ARG+                                  69                                    n2                                      -0.5000                                          394     MM11         13.576630592                21.822328796                       69.979103088                              ARG+                                  69                                    hn                                       0.3600                                          395     MM12         12.090744019                21.411947250                       70.936080933                              ARG+                                  69                                    hn                                       0.3600                                          396     MM2 12.989639282                22.056533813                       73.074882507                              ARG+                                  69                                    n2                                      -0.5000                                          397     MM21         12.033639908                21.696094513                       73.066261292                              ARG+                                  69                                    hn                                       0.3600                                          398     MM22         13.529273987                22.372974396                       73.883934021                              ARG+                                  69                                    hn                                       0.3600                                          399     N   19.436628342                26.928932190                       71.074501038                              ARG+                                  70                                    n -0.5000                                          400     CA  19.223009109                27.811206818                       69.878326416                              ARG+                                  70                                    ca                                       0.1200                                          401     MN  20.357131958                26.451332092                       71.165985107                              ARG+                                  70                                    hn                                       0.2800                                          402     MA  19.087514877                27.065124512                       69.071128845                              ARG+                                  70                                    h  0.1000                                          403     C   20.512538910                28.575149536                       69.398536682                              ARG+                                  70                                    c'                                       0.3800                                          404     O   20.872812271                28.468791962                       68.228363037                              ARG+                                  70                                    o'                                      -0.3800                                          405     CB  17.935552597                28.697717667                       69.732887268                              ARG+                                  70                                    c2                                      -0.2000                                          406     MB1 17.889257431                29.085472107                       68.694030762                              ARG+                                  70                                    h  0.1100                                          407     MB2 17.053388596                28.033058167                       69.807891846                              ARG+                                  70                                    h  0.1100                                          408     CG  17.775762558                29.883768082                       70.717842102                              ARG+                                  70                                    c2                                      -0.2000                                          409     MG1 18.004568100                29.946030045                       71.747383118                              ARG+                                  70                                    h  0.1300                                          410     MG2 18.540782928                30.651863098                       70.495643616                              ARG+                                  70                                    h  0.1300                                          411     CD  16.368116379                30.502414703                       70.693214417                              ARG+                                  70                                    c2                                      -0.0900                                          412     MD1 16.095691681                30.812221527                       69.664886475                              ARG+                                  70                                    h  0.1300                                          413     MD2 15.630161285                29.711160660                       70.937812805                              ARG+                                  70                                    h  0.1300                                          414     ME  16.255954742                31.590759277                       71.711013794                              ARG+                                  70                                    n1                                      -0.5000                                          415     ME  16.253637314                31.350564957                       72.706428528                              ARG+                                  70                                    hn                                       0.3600                                          416     CE  16.144437790                32.900745392                       71.464561462                              ARG+                                  70                                    cr                                       0.4500                                          417     MM1 16.055492401                33.712890625                       72.481109619                              ARG+                                  70                                    n2                                      -0.5000                                          418     MM11         16.071464539                33.294216156                       73.413330078                              ARG+                                  70                                    hn                                       0.3600                                          419     MM12         15.975571632                34.708621979                       72.277374268                              ARG+                                  70                                    hn                                       0.3600                                          420     MM2 16.120351791                33.420074463                       70.259872437                              ARG+                                  70                                    n2                                      -0.5000                                          421     MM21         16.025018692                34.432674408                       70.167800903                              ARG+                                  70                                    hn                                       0.3600                                          422     MM22         16.187112808                32.736862183                       69.505996704                              ARG+                                  70                                    hn                                       0.3600                                          423     N   21.115812302                29.562902451                       70.071769714                              TYRC                                  71                                    n -0.5000                                          424     MN  21.515977859                30.157514572                       69.338050842                              TYRC                                  71                                    hn                                       0.2800                                          425     CA  22.034273148                29.314456940                       71.218978882                              TYRC                                  71                                    ca                                       0.1200                                          426     MA  22.671009064                28.444923401                       70.976676941                              TYRC                                  71                                    h  0.1000                                          427     C   21.352920532                28.953493118                       72.563385010                              TYRC                                  71                                    c'                                       0.4100                                          428     OCT 20.392858505                29.553161621                       73.048652649                              TYRC                                  71                                    o'                                      -0.3800                                          429     O   21.928325653                27.853042603                       73.145797729                              TYRC                                  71                                    oh                                      -0.3800                                          430     MO  21.429273605                27.558662415                       73.909782410                              TYRC                                  71                                    ho                                       0.3500                                          431     CB  22.969152451                30.555358887                       71.361984253                              TYRC                                  71                                    c2                                      -0.2000                                          432     MB1 22.368267059                31.487627029                       71.355415344                              TYRC                                  71                                    h  0.1000                                          433     MB2 23.401992798                30.559690475                       72.381835938                              TYRC                                  71                                    h  0.1000                                          434     CG  24.144546509                30.666173935                       70.352111816                              TYRC                                  71                                    cp                                       0.0000                                          435     CD1 23.927837372                31.126276016                       69.044418335                              TYRC                                  71                                    cp                                      -0.1000                                          436     MD1 22.944635391                31.424867630                       68.717597961                              TYRC                                  71                                    h  0.1000                                          437     CE1 24.987041473                31.212265015                       68.143028259                              TYRC                                  71                                    cp                                      -0.1000                                          438     ME1 24.819047928                31.555503845                       67.131973267                              TYRC                                  71                                    h  0.1000                                          439     CZ  26.273118973                30.861675262                       68.542274475                              TYRC                                  71                                    cp                                       0.0300                                          440     OM  27.314481735                30.957365036                       67.652267456                              TYRC                                  71                                    oh                                      -0.3800                                          441     MN  26.980180740                31.236070633                       66.796859741                              TYRC                                  71                                    ho                                       0.3500                                          442     CE2 26.504697800                30.422637939                       69.841377258                              TYRC                                  71                                    cp                                      -0.1000                                          443     ME2 27.503232956                30.148866653                       70.140815735                              TYRC                                  71                                    h  0.1000                                          444     CD2 25.447391510                30.325349808                       70.743820190                              TYRC                                  71                                    cp                                      -0.1000                                          445     MD2 25.652494431                29.968608856                       71.745338440                              TYRC                                  71                                    h  0.1000                                          446     __________________________________________________________________________      *The numbering of the amino acids is shifted by minus 6 relative to the      sequence SEQ ID No. 1.

                                      TABLE NO. 2     __________________________________________________________________________                              residue                                    atom type,     Atom                     name and                                    charge and     name         x      y      z      no.   no.     __________________________________________________________________________     N   24.753730401                26.435615520                       68.246300842                              CYSn                                  40                                    nj                                      -0.5000                                          1     CA   24.503000259                26.356292725                       69.707687378                              CYSn                                  40                                    ca                                      0.1200                                          2     NN1 23.861560822                26.429992676                       67.734397888                              CYSn                                  40                                    hn                                      0.1400                                          3     NN2 25.250690460                25.603437424                       67.909477234                              CYSn                                  40                                    hn                                      0.1400                                          4     NA   23.890571594                27.247760773                       69.940788269                              CYSn                                  40                                    h 0.100                                          5     C     25.747190475                26.505004883                       70.632949829                              CYSn                                  40                                    c'                                      0.3800                                          6     O     25.611124039                27.204542160                       71.634971619                              CYSn                                  40                                    o'                                      -0.3800                                          7     CB   23.602088928                25.125436783                       69.979545593                              CYSn                                  40                                    c2                                      -0.3000                                          8     HB1 22.555475225                25.378625870                       69.716011047                              CYSn                                  40                                    h 0.1000                                          9     HB2 23.865217209                24.277284622                       69.317871094                              CYSn                                  40                                    h 0.1000                                          10     SG   23.613842010                24.466741562                       71.679084778                              CYSn                                  40                                    s1                                      0.1000                                          11     N     26.910152435                25.881416321                       70.350471497                              SER 41                                    n -0.5000                                          12     CA   28.052598953                25.721915930                       71.310394287                              SET 41                                    ca                                      0.1200                                          13     HN   26.921674728                25.473436356                       69.412322998                              SER 41                                    hn                                      0.2800                                          14     RA   27.761577606                24.886217117                       71.978584290                              SER 41                                    h 0.1000                                          15     C     28.477056503                26.929338455                       72.226699829                              SER 41                                    c'                                      0.3800                                          16     O     28.412267685                28.097608566                       71.829902649                              SER 41                                    o'                                      -0.3800                                          17     CB   28.257335662                25.212779999                       70.480110168                              SER 41                                    c2                                      -0.1700                                          18     KB1 28.957101822                24.401222229                       69.786743164                              SER 41                                    h 0.1000                                          19     KB2 29.657680511                26.030597687                       69.845893860                              SER 41                                    h 0.1000                                          20     CG   30.284814825                24.713315964                       71.339111328                              SER 41                                    ch                                      -0.3800                                          21     NG   31.027490616                24.448732376                       70.785003662                              SER 41                                    hc                                      0.3500                                          22     N     28.904022217                24.617259979                       73.466476440                              GLY 42                                    n -0.5000                                          23     CA   29.226711273                27.639400482                       74.497131348                              GLY 42                                    cg                                      0.0200                                          24     HX   29.140472412                25.626163483                       73.587532043                              GLY 42                                    hn                                      0.2800                                          25     KA1 28.567264557                28.519987106                       74.394309988                              GLY 42                                    h 0.1000                                          26     KA2 28.949186325                27.229663849                       75.483924866                              GLY 42                                    h 0.1000                                          27     C     30.728670120                28.040773392                       74.587509155                              GLY 42                                    c'                                      0.3800                                          28     O     31.522300720                27.171119690                       74.961143494                              GLY 42                                    o'                                      -0.3800                                          29     N     31.175983429                29.296203613                       74.282577515                              PRO 43                                    n -0.4200                                          30     CA   32.627410889                29.612504959                       74.140579224                              PRO 43                                    ca                                      0.0600                                          31     KA   33.153442383                28.711788635                       73.723632813                              PRO 43                                    h 0.1000                                          32     CO   30.295356750                30.374078751                       73.784042358                              PRO 43                                    c2                                      0.0600                                          33     KD1 29.605636597                30.735929489                       74.571166992                              PRO 43                                    h 0.1000                                          34     KD2 29.683467865                30.826647568                       72.928161621                              PRO 43                                    h 0.1000                                          35     C     33.389945984                30.112844467                       75.429031372                              PRO 43                                    c'                                      0.3800                                          36     O     32.754360199                30.681560516                       76.327354431                              PRO 43                                    o'                                      -0.3800                                          37     CB   32.565906525                30.710325241                       73.057388306                              PRO 43                                    c2                                      -0.2000                                          38     KB1 33.449081421                31.378034592                       73.055488586                              PRO 43                                    h 0.1000                                          39     KB2 32.524932861                30.247560501                       72.051818848                              PRO 43                                    h 0.1000                                          40     CG   31.263490677                31.466632843                       73.332626343                              PRO 43                                    c2                                      -0.2000                                          41     KG1 31.413110733                32.204200745                       74.146759033                              PRO 43                                    h 0.1000                                          42     KG2 30.894020081                32.022109985                       72.450286865                              PRO 43                                    h 0.1000                                          43     N     34.754848480                30.015562057                       75.523460388                              PRO 44                                    n -0.4200                                          44     CA   35.553565979                30.763086319                       76.536285400                              PRO 44                                    ca                                      0.0600                                          45     KA   35.083564758                30.695350647                       77.536567688                              PRO 44                                    h 0.1000                                          46     CD   35.574893951                29.225835648                       74.665214519                              PRO 44                                    c2                                      0.0600                                          47     KD1 35.471595764                29.373666763                       73.588935852                              PRO 44                                    h 0.1000                                          48     KD2 35.281650543                28.076414108                       74.807395935                              PRO 44                                    h 0.1000                                          49     C     35.767509460                32.265411377                       76.141113281                              PRO 44                                    c'                                      0.3300                                          50     O     36.544441223                32.599441528                       75.238464355                              PRO 44                                    c'                                      0.3800                                          51     CB   36.849227905                29.927103043                       76.567779541                              PRO 44                                    c2                                      -0.2000                                          52     HB1 37.732776642                30.502979279                       76.899009705                              PRO 44                                    h 0.1000                                          53     HB2 36.733722687                29.095364598                       72.253833069                              PRO 44                                    h 0.1000                                          54     CG   37.005489349                29.369808197                       75.152618408                              PRO 44                                    c2                                      -0.2000                                          55     MG1 37.502185822                30.119005203                       74.504158020                              PRO 44                                    h 0.1000                                          56     MG2 37.625408173                28.451385498                       75.115615845                              PRO 44                                    h 0.1000                                          57     N     35.047088623                33.173435211                       76.816978455                              ALA 45                                    n -0.5000                                          58     CA   35.011333466                34.609920502                       76.449218750                              ALA 45                                    ca                                      0.1200                                          59     MN   34.471405029                32.798248291                       77.590728760                              ALA 45                                    hn                                      0.2800                                          60     HA   35.065380096                34.699813843                       75.343650818                              ALA 45                                    h 0.1000                                          61     C     36.187728882                35.414947510                       77.090148926                              ALA 45                                    c'                                      0.3800                                          62     O     36.133388519                35.819305420                       78.255142212                              ALA 45                                    o'                                      -0.3800                                          63     CB   33.615478516                35.135440826                       76.831176758                              ALA 45                                    c3                                      -0.3000                                          64     HB1 33.490375519                36.187480927                       76.517280579                              ALA 45                                    h 0.1000                                          65     HB2 32.811222076                34.556259155                       76.338432312                              ALA 45                                    h 0.1000                                          66     HB3 33.433517456                35.098365784                       77.922439575                              ALA 45                                    h 0.1000                                          67     N     37.264499664                35.613868713                       76.306388855                              ALA 46                                    n -0.5000                                          68     HN   37.248832703                35.072978973                       75.433502197                              ALA 46                                    hn                                      0.2800                                          69     CA   38.503662109                36.298694611                       76.764076233                              ALA 46                                    ca                                      0.1200                                          70     MA   38.303600311                36.883266449                       77.688095093                              ALA 46                                    h 0.1000                                          71     C     39.082061768                37.273509979                       75.687866211                              ALA 46                                    c'                                      0.3800                                          72     O     38.951850891                37.052509308                       74.481193542                              ALA 46                                    c'                                      -0.3800                                          73     CB   39.509185791                35.179004669                       77.103065491                              ALA 46                                    c3                                      -0.3000                                          74     HB1 40.441535950                35.582756042                       77.525072327                              ALA 46                                    h 0.1000                                          75     HB2 39.106670380                34.460605621                       77.839447021                              ALA 46                                    h 0.1000                                          76     HB3 39.780502319                34.997728729                       76.205062866                              ALA 46                                    h 0.1000                                          77     N     39.768814087                38.344066620                       76.133750916                              ALA 47                                    n 0.5000                                          78     CA   40.322643280                39.391151428                       75.225708008                              ALA 47                                    ca                                      0.1200                                          79     MN   39.783836365                38.455593109                       77.149337769                              ALA 47                                    hn                                      0.2800                                          80     HA   39.932807922                39.265201569                       74.196365356                              ALA 47                                    h 0.1000                                          81     C     41.900882721                39.401574817                       75.126907349                              ALA 47                                    c'                                      0.3800                                          82     O     42.538444519                40.171451569                       75.854652405                              ALA 47                                    o'                                      -0.3800                                          83     CB   39.728843689                40.731719971                       75.714279175                              ALA 47                                    c3                                      -0.3000                                          84     HB1 40.043342590                41.567916870                       75.059875488                              ALA 47                                    h 0.1000                                          85     HB2 38.621978760                40.726497630                       75.711242676                              ALA 47                                    h 0.1000                                          86     HB3 40.062076569                40.987442017                       76.739311218                              ALA 47                                    h 0.1000                                          87     N     42.578651428                38.603999056                       74.242019653                              PRO 48                                    n -0.4200                                          88     CA   44.052674976                38.702857971                       74.013595581                              PRO 48                                    ca                                      0.0600                                          89     HA   44.576034546                38.850185394                       74.977058411                              PRO 48                                    h 0.1000                                          90     CO   41.956359863                37.474979401                       73.520225525                              PRO 48                                    c2                                      0.0600                                          91     HD1 41.114963531                37.378272858                       72.872795105                              PRO 48                                    h 0.1000                                          92     HD2 41.576156616                36.723918915                       74.239135742                              PRO 48                                    h 0.1000                                          93     C     44.492458344                39.820354462                       73.002609253                              PRO 48                                    c'                                      0.3800                                          94     O     43.782276154                40.131282806                       72.040626526                              PRO 48                                    o'                                      -0.3800                                          95     CB   44.356296539                37.288741516                       73.479736328                              PRO 48                                    c2                                      -0.2000                                          96     HB1 45.273612976                37.234390259                       72.865592957                              PRO 48                                    h 0.1000                                          97     HB2 44.813816833                36.591526031                       74.322021484                              PRO 48                                    h 0.1000                                          98     CG   43.102409363                36.884414673                       72.700988770                              PRO 48                                    c2                                      -0.2000                                          99     HG1 43.119277954                37.331111908                       71.685241699                              PRO 48                                    h 0.1000                                          100     HG2 43.010280609                35.788948059                       72.572326660                              PRO 48                                    h 0.1000                                          101     N     45.709655762                40.366821289                       73.185493469                              GLY 49                                    n -0.5000                                          102     CA   46.317604065                41.332912445                       72.214889526                              GLY 49                                    cg                                      0.0200                                          103     HN   46.169986725                40.089691162                       74.058357239                              GLY 49                                    hn                                      0.2800                                          104     HA1 45.654991150                41.537052153                       71.351181030                              GLY 49                                    h 0.1000                                          105     HA2 46.406318665                42.313266754                       72.719123840                              GLY 49                                    h 0.1000                                          106     C     47.710880280                40.963481903                       71.854037476                              GLY 49                                    c'                                      0.3800                                          107     O     48.630664825                41.772521973                       71.754951477                              GLY 49                                    o'                                      -0.3800                                          108     N     47.830738068                39.763301849                       71.063682556                              HIS 50                                    n -0.5000                                          109     HN   46.918842316                39.310573578                       70.943237305                              HIS 50                                    hn                                      0.2800                                          110     CA   49.045799255                39.210880280                       70.375061035                              HIS 50                                    ca                                      0.1200                                          111     HA   49.315334220                38.320884705                       70.972137451                              HIS 50                                    h 0.1000                                          112     C     50.433021545                39.981941223                       70.267852783                              HIS 50                                    c'                                      0.3800                                          113     O     50.773132324                40.456230164                       69.178672791                              HIS 50                                    o'                                      -0.3800                                          114     CB   28.558776855                32.590164165                       69.024101257                              HIS 50                                    c2                                      -0.2000                                          115     HB1 49.390335083                32.009521484                       68.577232361                              HIS 50                                    h 0.1000                                          116     HB2 47.792594910                37.817192078                       69.227330181                              HIS 50                                    h 0.1000                                          117     CG   47.997627258                39.545143127                       67.956726074                              HIS 50                                    c5                                      0.1000                                          118     HD1 46.669281006                39.956676482                       67.918121338                              HIS 50                                    np                                      -0.4200                                          119     CE1 46.730144501                40.789539337                       66.829002380                              HIS 50                                    c5                                      0.2700                                          120     NE2 47.911670685                40.950614929                       66.152328491                              HIS 50                                    np                                      -0.5000                                          121     CD2 48.729324341                40.126430511                       66.904235840                              HIS 50                                    c5                                      0.0100                                          122     HE1 45.843067169                41.327060699                       66.517631531                              HIS 50                                    h 0.1300                                          123     HE2 48.138290405                41.548683167                       65.349815369                              HIS 50                                    hn                                      0.2800                                          124     HD2 49.789726257                39.981491089                       66.738136292                              HIS 50                                    h 0.1300                                          125     H     51.307849884                40.071182251                       71.317932129                              PRO 51                                    n -0.4200                                          126     CA   52.692558289                40.596851349                       71.184913635                              PRO 51                                    ca                                      0.0600                                          127     HA   52.742668152                41.390510559                       70.412712097                              PRO 51                                    h 0.1000                                          128     CD   50.980678558                39.703777313                       72.706970215                              PRO 51                                    c2                                      0.0600                                          129     HD1 50.998199463                38.605384827                       72.818397522                              PRO 51                                    h 0.1000                                          130     HD2 49.987606049                40.071315765                       73.019950867                              PRO 51                                    h 0.1000                                          131     C     53.739063263                39.471630096                       70.880722046                              PRO 51                                    c'                                      0.3800                                          132     O     53.708900452                38.394466400                       71.488830566                              PRO 51                                    o'                                      -0.3800                                          133     CB   52.868911743                41.240936279                       72.572486877                              PRO 51                                    c2                                      -0.2000                                          134     HB1 53.929355621                41.364253998                       72.864852905                              PRO 51                                    h 0.1000                                          135     HB2 52.429229736                42.258647919                       72.565872192                              PRO 51                                    h 0.1000                                          136     CG   52.087848663                40.349472046                       73.547019958                              PRO 51                                    c2                                      -0.2000                                          137     HG1 52.750400543                39.566276550                       73.963310242                              PRO 51                                    h 0.1000                                          138     HG2 51.686916351                40.923263550                       74.403717041                              PRO 51                                    h 0.1000                                          139     H     54.676445007                39.726749420                       69.946899414                              LEU 52                                    n -0.5000                                          140     CA   55.768096924                38.764469147                       69.570259094                              LEU 52                                    ca                                      0.1200                                          141     HN   54.573589325                40.637012482                       69.488586426                              LEU 52                                    hn                                      0.2800                                          142     HA   56.414031982                39.325927734                       68.869346619                              LEU 52                                    h 0.1000                                          143     C     55.281269073                37.540004730                       68.718757629                              LEU 52                                    c'                                      0.3800                                          144     O     55.654800415                37.411125183                       67.550910950                              LEU 52                                    o'                                      -0.3800                                          145     CB   56.713882446                38.354763031                       70.751991272                              LEU 52                                    c2                                      -0.2000                                          146     HB1 56.125553131                37.737205505                       71.456863403                              LEU 52                                    h 0.1000                                          147     HB2 57.488136292                37.658962250                       70.374801636                              LEU 52                                    h 0.1000                                          148     CG   57.411731720                39.487998962                       71.552589417                              LEU 52                                    c1                                      -0.1000                                          149     HG   56.652648926                40.244640350                       71.834617615                              LEU 52                                    h 0.1000                                          150     CD1 58.010108948                38.936943054                       72.859535217                              LEU 52                                    c3                                      -0.3000                                          151     HD11         58.475826263                39.735752106                       73.467353821                              LEU 52                                    h 0.1000                                          152     HD12         57.236072540                38.469894409                       73.497505188                              LEU 52                                    h 0.1000                                          153     HD13         38.787303925                38.171112061                       72.675750732                              LEU 52                                    h 0.1000                                          154     CD2 58.517623901                40.187110901                       70.742630005                              LEU 52                                    c1                                      -0.3000                                          155     HD21         58.993679047                41.001243591                       71.321037292                              LEU 52                                    h 0.1000                                          156     HD22         59.321178436                39.487018585                       70.445312500                              LEU 52                                    h 0.1000                                          157     HD23         58.125030518                40.647811890                       69.818283081                              LEU 52                                    h 0.1000                                          158     H     54.475013733                36.643035889                       69.315246582                              ALA 53                                    n -0.5000                                          159     CA   53.896503448                35.451416016                       68.639259338                              ALA 53                                    ca                                      0.1200                                          160     HN   54.100524902                37.007503510                       70.205230713                              ALA 53                                    hn                                      0.2800                                          161     HA   53.553531647                35.773445129                       67.634338379                              ALA 53                                    h 0.1000                                          162     C     52.602260590                34.908508301                       69.353744507                              ALA 53                                    c'                                      0.3800                                          163     O     51.589202881                34.828308105                       68.650024414                              ALA 53                                    o'                                      -0.3800                                          164     CB   54.970031738                34.364234924                       68.394615173                              ALA 53                                    c3                                      -0.3000                                          165     HB1 54.534633636                33.449993134                       67.949813843                              ALA 53                                    h 0.1000                                          166     HB2 55.742454529                34.720954895                       67.688003540                              ALA 53                                    h 0.1000                                          167     HB3 55.500236511                34.068145752                       69.316299438                              ALA 53                                    h 0.1000                                          168     H     52.528438568                34.519321442                       70.670852661                              PRO 54                                    h -0.4200                                          169     CA   51.288402557                33.944232941                       71.268875122                              PRO 54                                    ca                                      0.0600                                          170     HA   50.860397339                33.194515228                       70.573081970                              PRO 54                                    h 0.1000                                          171     CD   53.678298950                34.513732910                       71.601814270                              PRO 54                                    c2                                      0.0600                                          172     HD1 54.146690369                35.509193420                       71.717323303                              PRO 54                                    h 0.1000                                          173     HD2 54.456859589                33.804187775                       71.264945984                              PRO 54                                    h 0.1000                                          174     C     50.163700104                34.973747253                       71.631500244                              PRO 54                                    c'                                      0.3800                                          175     O     50.381851136                36.186935425                       71.709472650                              PRO 54                                    o'                                      -0.3800                                          176     CB   51.868888855                33.209735870                       72.497810364                              PRO 54                                    c2                                      -0.2000                                          177     HB1 51.140216827                33.071922302                       73.319641113                              PRO 54                                    h 0.1000                                          178     HB2 52.201725006                32.193626404                       72.207275391                              PRO 43                                    h 0.1000                                          179     CG   53.074722290                34.047100067                       72.925216675                              PRO 54                                    c2                                      -0.2000                                          180     HG1 52.742275238                34.920612335                       73.520271301                              PRO 54                                    h 0.1000                                          181     HG2 53.794158936                33.482940674                       73.547462463                              PRO 54                                    h 0.1000                                          182     N     48.950717926                34.451953345                       71.882225037                              GLY 55                                    n -0.5000                                          183     CA   47.799011230                35.264900208                       72.354187012                              GLY 55                                    cg                                      0.2000                                          184     HN   48.913242340                33.427757262                       71.833122253                              GLY 55                                    hn                                      0.2800                                          185     HA1 47.829734802                36.291049957                       71.943481445                              GLY 55                                    h 0.1000                                          186     HA2 46.873668671                34.838825226                       71.925086975                              GLY 55                                    h 0.1000                                          187     C     47.624210358                35.262092590                       73.898422241                              GLY 55                                    c'                                      0.3800                                          188     O     47.184692383                34.228916168                       74.411125183                              GLY 55                                    o'                                      -0.3800                                          189     N     47.910079956                36.345748901                       74.679351807                              PRO 56                                    n -0.4200                                          190     CA   47.789894104                36.319248199                       76.162750244                              PRO 56                                    ca                                      0.0600                                          191     HA   48.177116394                35.361667633                       76.567420959                              PRO 56                                    h 0.1000                                          192     CD   48.602729797                37.547939301                       74.184982300                              PRO 56                                    c2                                      0.0600                                          193     HD1 48.046642303                38.065422058                       73.380996704                              PRO 56                                    h 0.1000                                          194     HD2 49.595470428                37.261718750                       73.794586182                              PRO 56                                    h 0.1000                                          195     C     46.326736450                36.529109955                       76.663719177                              PRO 56                                    c'                                      0.3800                                          196     O     45.857757568                37.657508850                       76.842826843                              PRO 56                                    o'                                      -0.3800                                          197     CB   48.777050018                37.430667877                       76.578651428                              PRO 56                                    c2                                      -0.2000                                          198     HB1 48.524734497                37.898471832                       77.549873352                              PRO 56                                    h 0.1000                                          199     HB2 49.795513153                37.009433746                       76.691307068                              PRO 56                                    h 0.1000                                          200     CG   48.752750397                38.431644440                       75.422836304                              PRO 56                                    c2                                      -0.2000                                          201     HG1 47.879917145                39.105598450                       75.522354126                              PRO 56                                    h 0.1000                                          202     HG2 49.655212402                39.069591522                       75.391311646                              PRO 56                                    h 0.1000                                          203     N     45.616943359                35.415058136                       76.897827148                              HIS 57                                    n -0.5000                                          204     HN   46.014194489                34.579135895                       76.450401306                              HIS 57                                    hn                                      0.2800                                          205     CA   44.212440491                35.434158325                       77.393867493                              HIS 57                                    ca                                      0.1200                                          206     HA   43.635601044                36.135776520                       76.762809753                              HIS 57                                    h 0.1000                                          207     C     44.146274567                35.882919312                       78.904235840                              HIS 57                                    c'                                      0.3800                                          208     O     44.718753815                35.174930573                       79.742973328                              HIS 57                                    o'                                      -0.3800                                          209     CB   43.577262878                34.025093079                       77.198188782                              HIS 57                                    c2                                      -0.2000                                          210     HB1 44.242053986                33.250709534                       77.626487732                              HIS 57                                    h 0.1000                                          211     HB2 42.665222168                33.964206696                       77.822631836                              HIS 57                                    h 0.1000                                          212     CG   43.190551758                33.606594086                       75.770996094                              HIS 57                                    c5                                      0.1000                                          213     HD1 44.009815216                33.724720001                       74.654800415                              HIS 57                                    np                                      -0.4200                                          214     CE1 43.220783234                33.103317261                       73.724327087                              HIS 57                                    c5                                      0.2700                                          215     HE1 42.000507355                32.603866577                       74.087806702                              HIS 57                                    np                                      -0.5000                                          216     CD2 42.008693695                32.921348572                       75.433784485                              HIS 57                                    c5                                      0.0100                                          217     HE1 43.579235077                32.995296478                       72.708923340                              HIS 57                                    h 0.1300                                          218     HE2 41.324546814                32.061363220                       73.538429260                              HIS 57                                    hn                                      0.2800                                          219     HD2 41.238662720                32.638732910                       76.138023376                              HIS 57                                    h 0.1300                                          220     N     43.493415833                37.014365150                       79.314620972                              PRO 58                                    n -0.4200                                          221     CA   43.576023102                37.521991730                       80.713310242                              PRO 58                                    ca                                      0.0.600                                          222     HA   44.640773772                37.575522258                       81.019523621                              PRO 58                                    h 0.1000                                          233     CD   42.828823090                37.960189819                       78.395561218                              PRO 58                                    c2                                      0.0600                                          224     HD1 42.088195801                37.471439362                       77.735382080                              PRO 58                                    h 0.1000                                          225     HD2 43.573791504                38.467678070                       77.749885559                              PRO 58                                    h 0.1000                                          226     C     42.782455444                36.660888672                       81.747703552                              PRO 58                                    c'                                      0.3800                                          227     O     41.546211243                36.662284851                       81.766304016                              PRO 58                                    o'                                      -0.3800                                          228     CB   43.067096710                38.972381592                       80.563407898                              PRO 58                                    c2                                      -0.2000                                          229     HB1 42.553604126                39.355644226                       81.465919495                              PRO 58                                    h 0.1000                                          230     HB2 43.922630310                39.652961731                       80.382720947                              PRO 58                                    h 0.1000                                          231     CG   42.156223297                38.958541870                       79.333923340                              PRO 58                                    c2                                      -0.2000                                          232     HG1 41.146640778                38.599040985                       79.611282349                              PRO 58                                    h 0.1000                                          233     HG2 42.036239624                39.956802368                       78.871780396                              PRO 58                                    h 0.1000                                          234     N     43.511478424                35.934528351                       82.617271423                              ALA 59                                    n -0.5000                                          235     HN   44.509765625                35.905010233                       82.386589050                              ALA 59                                    hn                                      0.2800                                          236     CA   42.913761139                34.928112030                       83.544029236                              ALA 59                                    ca                                      0.1200                                          237     HA   42.154701233                34.341350555                       82.987930298                              ALA 59                                    h 0.1000                                          238     C     42.181308746                35.481468201                       84.821502686                              ALA 59                                    c'                                      0.3800                                          239     O     42.459964752                35.112228394                       85.965652466                              ALA 59                                    o'                                      -0.3800                                          240     CB   44.063701630                33.948829651                       83.858291626                              ALA 59                                    c3                                      -0.3000                                          241     HB1 43.708641052                33.104522705                       84.478195190                              ALA 59                                    h 0.1000                                          242     HB2 44.502014160                33.503505707                       82.944122314                              ALA 59                                    h 0.1000                                          243     HB3 44.882900238                34.433902740                       84.422813416                              ALA 59                                    h 0.1000                                          244     N     41.178901672                36.333106995                       84.577079773                              ALA 60                                    n -0.5000                                          245     HN   41.112052917                36.562049866                       83.573013306                              ALA 60                                    hn                                      0.2800                                          246     CA   40.189502716                36.803413391                       85.578857422                              ALA 60                                    ca                                      0.1200                                          247     HA   40.008514404                35.981742859                       86.302818298                              ALA 60                                    h 0.1000                                          248     C     38.811019897                37.037807465                       84.860046387                              ALA 60                                    c'                                      0.3800                                          249     O     37.881835938                36.301105499                       85.205276489                              ALA 60                                    o'                                      -0.3800                                          250     CB   40.746566772                37.985549927                       86.401313782                              ALA 60                                    c3                                      -0.3000                                          251     HB1 39.997776031                38.372333527                       87.116531372                              ALA 60                                    h 0.1000                                          252     HB2 41.624504089                37.670467377                       86.996170044                              ALA 60                                    h 0.1000                                          253     HB3 41.079444885                38.830841064                       85.774902344                              ALA 60                                    h 0.1000                                          254     N     38.609931946                37.922218323                       83.826889038                              PRO 61                                    n -0.4200                                          255     CA   37.398490906                37.873073578                       82.950836182                              PRO 61                                    ca                                      0.0600                                          256     HA   36.485267638                37.839759827                       83.576164246                              PRO 61                                    h 0.1000                                          257     CD   39.552070618                39.001556396                       83.460945129                              PRO 61                                    c2                                      0.0600                                          258     HD1 40.590957642                38.653537750                       83.311111450                              PRO 61                                    h 0.1000                                          259     HD2 39.564567512                39.780746460                       84.247795105                              PRO 61                                    h 0.1000                                          260     C     37.256782532                36.700027466                       81.909835815                              PRO 61                                    c'                                      0.3800                                          261     O     36.243316650                36.657413483                       81.209106445                              PRO 61                                    o'                                      -0.3800                                          262     CB   37.477272034                39.256717682                       82.271400452                              PRO 61                                    c2                                      -0.2000                                          263     HB1 36.964195251                39.298637390                       81.290786743                              PRO 61                                    h 0.1000                                          264     HB2 36.981357574                40.016944885                       82.906776428                              PRO 61                                    h 0.1000                                          265     CG   38.972518921                39.562160492                       82.163795471                              PRO 61                                    c2                                      -0.2000                                          266     HG1 39.409160614                39.034259796                       81.293624878                              PRO 61                                    h 0.1000                                          267     HG2 39.183399200                40.640045166                       82.032295227                              PRO 61                                    h 0.1000                                          268     N     38.213741302                35.753643036                       81.804443359                              SER 62                                    n -0.5000                                          269     CA   38.144962311                34.600208282                       80.863403320                              SER 62                                    ca                                      0.1200                                          270     HN   39.009967804                35.895751953                       82.434890747                              SER 62                                    hn                                      0.2800                                          271     HA   37.892318726                34.983673096                       79.856529236                              SER 62                                    h 0.1000                                          272     C     37.085021973                31.524326324                       81.273231506                              SER 62                                    c'                                      0.3800                                          273     O     37.382484436                32.625560760                       82.070343018                              SER 62                                    o'                                      -0.3800                                          274     CB   39.569152832                33.994293213                       80.760513306                              SER 62                                    c2                                      -0.1700                                          275     HB1 39.601100922                33.242229462                       79.949050903                              SER 62                                    h 0.1000                                          276     HB2 40.217050934                34.759819031                       80.475799561                              SER 62                                    h 0.1000                                          277     CG   39.958038330                33.367904663                       81.986091614                              SER 62                                    oh                                      0.1000                                          278     HG   39.157264709                32.928077698                       82.316406250                              SER 62                                    ho                                      0.3500                                          279     N     35.853912254                33.643447876                       80.748901367                              SER 63                                    n -0.5000                                          280     CA   34.681579590                32.893772125                       81.280097961                              SER 63                                    ca                                      0.1200                                          281     HN   35.734226227                34.507610321                       80.200576782                              SER 63                                    hn                                      0.2800                                          282     HA   34.978878021                32.281963348                       82.158424377                              SER 63                                    h 0.1000                                          283     C     34.028987885                31.963005066                       80.214485168                              SER 63                                    c'                                      0.3800                                          284     O     33.624385834                32.404701233                       79.134857178                              SER 63                                    o'                                      -0.3800                                          285     CB   33.674541473                33.937458038                       81.815757751                              SER 63                                    c2                                      -0.1700                                          286     HB1 34.172107697                34.614356995                       82.538948059                              SER 63                                    h 0.1000                                          287     HB2 33.303077698                34.594402313                       81.003784180                              SER 63                                    h 0.1000                                          288     CG   32.576236725                33.301517487                       82.471984863                              SER 63                                    oh                                      -0.3800                                          289     HG   32.084590912                32.806625366                       81.807708740                              SER 63                                    ho                                      0.3500                                          290     N     33.852622986                30.677625656                       80.556045532                              TRP 64                                    n -0.5000                                          291     CA   33.070865631                29.709415436                       79.732810974                              TRP 64                                    ca                                      0.1200                                          292     HN   34.153343201                30.435497284                       81.506057739                              TRP 64                                    hn                                      0.2800                                          293     HA   33.375720978                29.840501785                       78.676765442                              TRP 64                                    h 0.1000                                          294     C     31.526346207                29.958730698                       79.816452026                              TRP 64                                    c'                                      0.3800                                          295     O     30.936735153                29.911306381                       80.900970459                              TRP 64                                    o'                                      -0.3800                                          296     CB   33.498836517                28.253648758                       80.086425781                              TRP 64                                    c2                                      -0.2000                                          297     HB1 32.936897276                27.550512314                       79.442245483                              TRP 64                                    h 0.1000                                          298     HB2 34.548812866                28.110004426                       79.767990112                              TRP 64                                    h 0.1000                                          299     CG   33.372638702                27.788063049                       81.551361084                              TRP 64                                    c5                                      0.0000                                          300     CD1 32.236022969                27.198472977                       82.145393372                              TRP 64                                    c5                                      0.0100                                          301     HE1 32.442039490                26.926628113                       83.513259888                              TRP 64                                    np                                      -0.5000                                          302     CE2 33.734771729                27.365074158                       83.750877380                              TRP 64                                    c5                                      0.1100                                          303     CD2 34.313194275                27.885219574                       82.565856934                              TRP 64                                    c5                                      0.0000                                          304     HD1 31.308589935                27.011001587                       81.622375488                              TRP 64                                    h 0.1000                                          305     HE1 31.781488419                26.532098770                       84.192108154                              TRP 64                                    hn                                      0.2800                                          306     CE3 35.633460999                28.410655975                       82.581642151                              TRP 64                                    cp                                      -0.1000                                          307     HE3 36.086208344                28.812973022                       81.686943054                              TRP 64                                    h 0.1000                                          308     CZ3 36.338203430                28.401332855                       83.786201477                              TRP 64                                    cp                                      -0.1000                                          309     HZ3 37.342418671                28.800062180                       83.816780090                              TRP 64                                    h 0.1000                                          310     CH2 35.766292572                27.887487411                       84.956939697                              TRP 64                                    cp                                      -0.1000                                          311     HH2 36.336753845                27.895225525                       85.875114441                              TRP 64                                    h 0.1000                                          312     CZ2 34.469055176                27.367534637                       84.959693909                              TRP 64                                    cp                                      -0.1000                                          313     HZ2 34.033626556                26.978828430                       85.868453979                              TRP 64                                    h 0.1000                                          314     N     30.884126663                30.253189087                       78.672058105                              GLY 65                                    n -0.5000                                          315     CA   29.433704376                30.522933426                       78.625785828                              GLY 54                                    cg                                      0.0200                                          316     HN   31.490442276                30.324731827                       77.837860107                              GLY 65                                    hn                                      0.2800                                          317     HA1 29.049486160                30.927183151                       79.604759216                              GLY 65                                    h 0.1000                                          318     HA2 29.301883698                31.462034225                       77.967575073                              GLY 65                                    h 0.1000                                          319     C     28.566276550                29.436250687                       78.049354553                              GLY 65                                    c'                                      0.3800                                          320     O     28.476503372                29.383417130                       76.820671082                              GLY 65                                    o'                                      -0.3800                                          321     N     27.919076920                28.520057678                       78.830680847                              PRO 66                                    n -0.4200                                          322     CA   27.254581451                27.307062149                       78.266265869                              PRO 66                                    ca                                      0.0600                                          323     HA   28.007204056                26.749544144                       77.674018860                              PRO 66                                    h 0.1000                                          324     CD   27.931732178                28.547025681                       80.307373047                              PRO 66                                    c2                                      0.0600                                          325     MD1 27.674114227                29.536535263                       80.727989197                              PRO 66                                    h 0.1000                                          326     MD2 28.930458069                28.264209747                       80.693565369                              PRO 66                                    h 0.1000                                          327     C     25.991449356                27.540506363                       77.367637634                              PRO 66                                    c'                                      0.3800                                          328     O     25.234470367                28.498056412                       77.550445557                              PRO 66                                    o'                                      -0.3800                                          329     CB   26.947065353                26.487516403                       79.540168762                              PRO 66                                    c2                                      -0.2000                                          330     HB1 26.021696091                25.883453369                       79.667781067                              PRO 66                                    h 0.1000                                          331     HB2 27.765922546                25.768106561                       79.730659485                              PRO 66                                    h 0.1000                                          332     CG   26.879997253                27.505201340                       80.680580139                              PRO 66                                    c2                                      -0.2000                                          333     HG1 25.878761292                27.974497401                       80.712356567                              PRO 66                                    h 0.1000                                          334     HG2 27.057765961                27.053478241                       81.674545288                              PRO 66                                    h 0.1000                                          335     N     25.764133453                26.624628067                       76.406608582                              CYS 67                                    n -0.5000                                          336     CA   24.578670502                26.665664673                       75.512832642                              CYS 67                                    ca                                      0.1200                                          337     HN   26.474828720                25.893449783                       76.320098877                              CYS 67                                    hn                                      0.2800                                          338     HA   24.437746048                27.701400757                       75.152099609                              CYS 67                                    h 0.1000                                          339     C     23.256376266                26.130277634                       76.174392700                              CYS 67                                    c'                                      0.3800                                          340     O     23.219629288                24.940492630                       76.516906738                              CYS 67                                    o'                                      -0.3800                                          341     CB   24.900175095                25.819908432                       74.260444641                              CYS 67                                    c2                                      -0.3000                                          342     HB1 25.807971954                26.178848267                       73.749794006                              CYS 67                                    h 0.1000                                          343     HB2 25.105169296                24.761932373                       74.532623291                              CYS 67                                    h 0.1000                                          344     CG   23.472158432                25.844451904                       73.133270264                              CYS 67                                    s1                                      0.1000                                          345     N     22.124137878                26.895683289                       76.264381409                              PRO 68                                    n -0.4200                                          346     CA   20.786550522                26.297697067                       76.829830933                              PRO 69                                    ca                                      0.0600                                          347     HA   20.877141953                25.506265650                       77.300582886                              PRO 68                                    h 0.1000                                          348     CD   22.161409378                28.364057541                       76.432929993                              PRO 68                                    c2                                      0.0600                                          349     HD1 22.628620148                28.901371002                       75.585960388                              PRO 68                                    h 0.1000                                          350     HD2 22.732339859                28.631174088                       77.345329285                              PRO 68                                    h 0.1000                                          351     C     20.190311432                25.593938828                       75.255737305                              PRO 68                                    c'                                      0.3800                                          352     O     20.566764832                24.451507568                       74.984413147                              PRO 68                                    o'                                      -0.3800                                          353     CB   20.033475876                27.483673096                       77.173645020                              PRO 68                                    c2                                      -0.2000                                          354     HB1 18.940057755                27.449979782                       77.017181396                              PRO 68                                    h 0.1000                                          355     HB2 20.183664322                27.458950043                       78.271354675                              PRO 68                                    h 0.1000                                          356     CG   20.687761307                28.746078491                       76.604209900                              PRO 68                                    c2                                      -0.2000                                          357     HG1 20.234010696                29.017192841                       75.632743835                              PRO 68                                    h 0.1000                                          358     HG2 20.559530258                29.623554230                       77.265640259                              PRO 68                                    h 0.1000                                          359     N     19.297069550                26.226366043                       74.460205078                              ARG+                                  69                                    n -0.5000                                          360     CA   18.727945328                25.594141006                       73.229873657                              ARG+                                  69                                    ca                                      0.1200                                          361     HN   18.899488449                27.074655533                       74.874603271                              ARG+                                  69                                    hn                                      0.2800                                          362     HA   19.468439102                24.889890671                       72.798027039                              ARG+                                  69                                    h 0.1000                                          363     C     18.426181793                26.666866302                       72.127845764                              ARG+                                  69                                    c'                                      0.3800                                          364     O     17.302417755                27.170328140                       72.057907104                              ARG+                                  69                                    o'                                      -0.3800                                          365     CB   17.487716675                24.741283417                       73.645401001                              ARG+                                  69                                    c2                                      -0.2000                                          366     HB1 17.790594101                24.038330078                       74.447227478                              ARG+                                  69                                    h 0.1100                                          367     HB2 16.742151260                25.409061432                       74.119949341                              ARG+                                  69                                    h 0.1100                                          368     CG   16.806570053                23.930654526                       72.510940552                              ARG+                                  69                                    c2                                      -0.2000                                          369     HG1 16.500089645                24.624885559                       71.702163696                              ARG+                                  69                                    h 0.1300                                          370     HG2 17.539186478                23.235509872                       72.053100586                              ARG+                                  69                                    h 0.1300                                          371     CD   15.574314117                23.148860931                       73.007453918                              ARG+                                  69                                    c2                                      -0.0900                                          372     HD1 15.890284538                22.374292374                       73.738624573                              ARG+                                  69                                    h 0.1300                                          373     HD2 14.902976036                23.843069077                       73.554016113                              ARG+                                  69                                    h 0.1300                                          374     HZ   14.865127563                22.521680832                       71.855873108                              ARG+                                  69                                    n1                                      -0.5000                                          375     HE   15.293711662                22.507183075                       70.926025391                              ARG+                                  69                                    hn                                      0.3600                                          376     CZ   13.645489693                21.980854034                       71.902374268                              ARG+                                  69                                    cr                                      0.4500                                          377     NH1 13.127552986                21.522832870                       70.798370361                              ARG+                                  69                                    n2                                      -0.5000                                          378     NH11         13.689088821                21.608518600                       69.948852539                              ARG+                                  69                                    hn                                      0.3600                                          379     NH12         12.188611031                21.122539520                       70.853851318                              ARG+                                  69                                    hn                                      0.3600                                          380     HN2 12.936479568                21.886768341                       73.000465393                              ARG+                                  69                                    n2                                      -0.500                                          381     NH21         12.008401871                21.462900162                       72.952354431                              ARG+                                  69                                    hn                                      0.3600                                          382     NH22         13.405644417                22.251142502                       73.831558228                              ARG+                                  69                                    hn                                      0.3600                                          383     N     19.430337904                26.966600418                       71.273384094                              ARG+                                  70                                    n -0.5000                                          384     CA   19.316965103                27.784936905                       70.016807556                              ARG+                                  70                                    ca                                      0.1200                                          385     HW   20.317523956                26.522159576                       71.527793884                              ARG+                                  70                                    hn                                      0.2800                                          386     HA   19.139877319                27.013025284                       69.241798401                              ARG+                                  70                                    h 0.1000                                          387     C     20.690151215                28.397680283                       69.573341370                              ARG+                                  70                                    c'                                      0.3800                                          388     O     21.267679214                27.963806152                       68.579154968                              ARG+                                  70                                    o'                                      -0.3800                                          389     CB   18.103752136                28.753881454                       69.796676636                              ARG+                                  70                                    c2                                      -0.2000                                          390     HB1 18.134477615                29.133897781                       68.754875183                              ARG+                                  70                                    h 0.1100                                          391     HB2 17.183977127                28.135446548                       69.816154480                              ARG+                                  70                                    h 0.1100                                          392     CG   17.944366455                29.952959061                       70.767364502                              ARG+                                  70                                    c2                                      -0.2000                                          393     HG1 18.201717377                29.630409241                       71.795295715                              ARG+                                  70                                    h 0.1300                                          394     HG2 18.686578751                30.737569809                       70.523498535                              ARG+                                  70                                    h 0.1300                                          395     CD   16.516407013                30.528032303                       70.757499695                              ARG+                                  70                                    c2                                      -0.0900                                          396     HD1 16.205293655                30.812314987                       69.732498169                              ARG+                                  70                                    h 0.1300                                          397     HD2 15.804359436                29.724859238                       71.041206360                              ARG+                                  70                                    h 0.1300                                          398     HE   16.396289825                31.632083883                       71.751823425                              ARG+                                  70                                    n1                                      -0.5000                                          399     NE   16.381929398                31.418577194                       72.754409790                              ARG+                                  70                                    hn                                      0.3600                                          400     CE   16.270032883                32.931114197                       71.475357056                              ARG+                                  70                                    cr                                      0.4500                                          401     HN1 16.119342804                33.758121490                       72.470024109                              ARG+                                  70                                    n2                                      -0.5000                                          402     HN11         16.097789764                33.352241516                       73.407394409                              ARG+                                  70                                    hn                                      0.3600                                          403     HN12         16.020824432                34.751869202                       72.242614746                              ARG+                                  70                                    hn                                      0.3600                                          404     NH2 16.295974731                33.427280426                       70.262794495                              ARG+                                  70                                    n2                                      -0.5000                                          405     HN21         16.191591263                34.436088562                       70.142570496                              ARG+                                  70                                    hn                                      0.3600                                          406     HN22         16.439620972                32.732715607                       69.527351379                              ARG+                                  70                                    hn                                      0.3600                                          407     N     21.215826035                29.506198883                       70.104598999                              TYRC                                  71                                    n -0.5000                                          408     HN   21.692993164                29.961484909                       69.319816589                              TYRC                                  71                                    hn                                      0.2800                                          409     CA   22.037544250                29.469150543                       71.348726365                              TYRC                                  71                                    ca                                      0.1300                                          410     HA   22.727062225                28.601654053                       71.296867371                              TYRC                                  71                                    h 0.1000                                          411     C     21.230804443                29.295030594                       72.663772583                              TYRC                                  71                                    c'                                      0.4100                                          412     CKT 20.420524597                30.105148315                       73.113685608                              TYRC                                  71                                    o'                                      -0.3800                                          413     O     21.522385052                28.107995987                       73.273979187                              TYRC                                  71                                    ch                                      -0.3800                                          414     HO   20.994127274                28.000011444                       74.066841125                              TYRC                                  71                                    ho                                      0.3500                                          415     CB   22.938613892                30.740638733                       71.402084351                              TYRC                                  71                                    c2                                      -0.2000                                          416     HB1 22.321226120                31.652799606                       71.283157349                              TYRC                                  71                                    h 0.1000                                          417     HB2 23.363500595                30.853305817                       72.420455933                              TYRC                                  71                                    h 0.1000                                          418     CG   24.110603333                30.760416031                       70.402580261                              TYRC                                  71                                    cp                                      0.000                                          419     CD1 23.933057785                31.274461746                       69.111869812                              TYRC                                  71                                    cp                                      -0.1000                                          420     HD1 22.977622986                31.679159164                       68.809974670                              TYRC                                  71                                    h 0.1000                                          421     CE1 24.985538483                31.264329910                       68.201301575                              TYRC                                  71                                    cp                                      -0.1000                                          422     HE1 24.833002090                31.650396347                       67.203536987                              TYRC                                  71                                    h 0.1000                                          423     CZ   26.227394104                30.757091522                       68.577186584                              TYRC                                  71                                    cp                                      0.0300                                          424     OH   27.265848160                30.762763977                       67.686424255                              TYRC                                  71                                    oh                                      -0.3800                                          425     HN   29.966634750                31.154380798                       66.863937378                              TYRC                                  71                                    ho                                      0.3500                                          426     CE2 26.415199280                30.251981735                       69.859985352                              TYRC                                  71                                    cp                                      -0.1000                                          427     HE2 27.377700806                29.852491379                       70.147377014                              TYRC                                  71                                    h 0.1000                                          428     CD2 25.360828400                30.253871918                       70.770927429                              TYRC                                  71                                    cp                                      -0.1000                                          429     HD2 25.521846771                29.846044540                       71.760574341                              TYRC                                  71                                    h 0.1000                                          430     __________________________________________________________________________      *The numbering of the amino acids is shifted by minus 6 relative to the      sequence SEQ ID No. 1

    __________________________________________________________________________     #             SEQUENCE LISTING     - (1) GENERAL INFORMATION:     -    (iii) NUMBER OF SEQUENCES: 8     - (2) INFORMATION FOR SEQ ID NO:1:     -      (i) SEQUENCE CHARACTERISTICS:     #acids    (A) LENGTH: 476 amino               (B) TYPE: amino acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: protein     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:     - Leu Gln Pro Gly Ala Glu Val Pro Val Val Tr - #p Ala Gln Glu Gly Ala     #                15     - Pro Ala Gln Leu Pro Cys Ser Pro Thr Ile Pr - #o Leu Gln Asp Leu Ser     #            30     - Leu Leu Arg Arg Ala Gly Val Thr Trp Gln Hi - #s Gln Pro Asp Ser Gly     #        45     - Pro Pro Ala Ala Ala Pro Gly His Pro Leu Al - #a Pro Gly Pro His Pro     #    60     - Ala Ala Pro Ser Ser Trp Gly Pro Arg Pro Ar - #g Arg Tyr Thr Val Leu     #80     - Ser Val Gly Pro Gly Gly Leu Arg Ser Gly Ar - #g Leu Pro Leu Gln Pro     #                95     - Arg Val Gln Leu Asp Glu Arg Gly Arg Gln Ar - #g Gly Asp Phe Ser Leu     #           110     - Trp Leu Arg Pro Ala Arg Arg Ala Asp Ala Gl - #y Glu Tyr Arg Ala Ala     #       125     - Val His Leu Arg Asp Arg Ala Leu Ser Cys Ar - #g Leu Arg Leu Arg Leu     #   140     - Gly Gln Ala Ser Met Thr Ala Ser Pro Pro Gl - #y Ser Leu Arg Ala Ser     145                 1 - #50                 1 - #55                 1 -     #60     - Asp Trp Val Ile Leu Asn Cys Ser Phe Ser Ar - #g Pro Asp Arg Pro Ala     #               175     - Ser Val His Trp Phe Arg Asn Arg Gly Gln Gl - #y Arg Val Pro Val Arg     #           190     - Glu Ser Pro His His His Leu Ala Glu Ser Ph - #e Leu Phe Leu Pro Gln     #       205     - Val Ser Pro Met Asp Ser Gly Pro Trp Gly Cy - #s Ile Leu Thr Tyr Arg     #   220     - Asp Gly Phe Asn Val Ser Ile Met Tyr Asn Le - #u Thr Val Leu Gly Leu     225                 2 - #30                 2 - #35                 2 -     #40     - Glu Pro Pro Thr Pro Leu Thr Val Tyr Ala Gl - #y Ala Gly Ser Arg Val     #               255     - Gly Leu Pro Cys Arg Leu Pro Ala Gly Val Gl - #y Thr Arg Ser Phe Leu     #           270     - Thr Ala Lys Trp Thr Pro Pro Gly Gly Gly Pr - #o Asp Leu Leu Val Thr     #       285     - Gly Asp Asn Gly Asp Phe Thr Leu Arg Leu Gl - #u Asp Val Ser Gln Ala     #   300     - Gln Ala Gly Thr Tyr Thr Cys His Ile His Le - #u Gln Glu Gln Gln Leu     305                 3 - #10                 3 - #15                 3 -     #20     - Asn Ala Thr Val Thr Leu Ala Ile Ile Thr Va - #l Thr Pro Lys Ser Phe     #               335     - Gly Ser Pro Gly Ser Leu Gly Lys Leu Leu Cy - #s Glu Val Thr Pro Val     #           350     - Ser Gly Gln Glu Arg Phe Val Trp Ser Ser Le - #u Asp Thr Pro Ser Gln     #       365     - Arg Ser Phe Ser Gly Pro Trp Leu Glu Ala Gl - #n Glu Ala Gln Leu Leu     #   380     - Ser Gln Pro Trp Gln Cys Gln Leu Tyr Gln Gl - #y Glu Arg Leu Leu Gly     385                 3 - #90                 3 - #95                 4 -     #00     - Ala Ala Val Tyr Phe Thr Glu Leu Ser Ser Pr - #o Gly Ala Gln Arg Ser     #               415     - Gly Arg Ala Pro Gly Ala Leu Pro Ala Gly Hi - #s Leu Leu Leu Phe Leu     #           430     - Thr Leu Gly Val Leu Ser Leu Leu Leu Leu Va - #l Thr Gly Ala Phe Gly     #       445     - Phe His Leu Trp Arg Arg Gln Trp Arg Pro Ar - #g Arg Phe Ser Ala Leu     #   460     - Glu Gln Gly Ile His Pro Arg Arg Leu Arg Al - #a Arg     465                 4 - #70                 4 - #75     - (2) INFORMATION FOR SEQ ID NO:2:     -      (i) SEQUENCE CHARACTERISTICS:     #pairs    (A) LENGTH: 32 base               (B) TYPE: nucleic acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: cDNA     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:     #          32      ACCA TAGGAGAGAT GT     - (2) INFORMATION FOR SEQ ID NO:3:     -      (i) SEQUENCE CHARACTERISTICS:     #pairs    (A) LENGTH: 40 base               (B) TYPE: nucleic acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: cDNA     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:     #    40            ACCC AGAACAGTGA GGTTATACAT     - (2) INFORMATION FOR SEQ ID NO:4:     -      (i) SEQUENCE CHARACTERISTICS:     #pairs    (A) LENGTH: 34 base               (B) TYPE: nucleic acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: cDNA     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:     #        34        GCTA GACAGCTCTG TGAA     - (2) INFORMATION FOR SEQ ID NO:5:     -      (i) SEQUENCE CHARACTERISTICS:     #pairs    (A) LENGTH: 37 base               (B) TYPE: nucleic acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: cDNA     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:     #      37          CCAG ACCATAGGAG AGATGTG     - (2) INFORMATION FOR SEQ ID NO:6:     -      (i) SEQUENCE CHARACTERISTICS:     #pairs    (A) LENGTH: 37 base               (B) TYPE: nucleic acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: cDNA     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:     #      37          CGAG GCCTGGCCCA GGCGCAG     - (2) INFORMATION FOR SEQ ID NO:7:     -      (i) SEQUENCE CHARACTERISTICS:     #pairs    (A) LENGTH: 35 base               (B) TYPE: nucleic acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: cDNA     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:     #       35         CAGA ACAGTGAGGT TATAC     - (2) INFORMATION FOR SEQ ID NO:8:     -      (i) SEQUENCE CHARACTERISTICS:     #pairs    (A) LENGTH: 35 base               (B) TYPE: nucleic acid               (C) STRANDEDNESS: single               (D) TOPOLOGY: linear     -     (ii) MOLECULE TYPE: cDNA     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:     #       35         TGGG CTAGACAGCT CTGTG     __________________________________________________________________________ 

We claim:
 1. A soluble polypeptide fraction consisting of at least one of the 4 immunoglobulin type extra-cellular domains of the LAG-3 protein (amino acids 1 to 4, 5 to 239, 240 to 330 and 331 to 412 of sequence SEQ ID NO:1), wherein one or more arginine (Arg) residues at positions 73, 75 and 76 of SEQ ID NO:1 are substituted with glutamic acid (Glu) and said at least one extra-cellular domain of LAG-3 protein is optionally fused to a supplementary peptide sequence as a fusion protein.
 2. A soluble polypeptide fraction according to claim 1, further bound to a toxin or a radioisotope.
 3. A soluble polypeptide fraction according to claim 1, further bound to a toxin or a radioisotope wherein said supplementary peptide sequence is present and comprises a portion of an immunoglobulin.
 4. A soluble polypeptide fraction according to claim 3, wherein said supplementary peptide sequence comprises a portion of an immunoglobulin of IgG4 isotype.
 5. A method for producing the soluble polypeptide fraction of claim 1, which soluble polypeptide fraction further comprises a portion of an immunoglobulin, comprising the steps of:inserting a DNA molecule comprising a fusion of fragments of cDNA coding for the polypeptide regions corresponding to LAG-3 or derived from LAG3with cDNA coding for the portion of the immunoglobulin; transfecting the DNA molecule into a host expression system; and producing the soluble polypeptide fraction by expression in the host.
 6. A soluble polypeptide fraction consisting of at least one of four immunoglobulin-type extracellular domains of LAG-3 protein corresponding to amino acid residues 1-4, 5-239, 240-330, and 331-412 of SEQ ID NO:1, fused to a supplementary peptide sequence as a fusion protein.
 7. A soluble polypeptide fraction according to claim 6, further bound to a toxin or a radioisotope.
 8. A soluble polypeptide fraction according to claim 6, further bound to a toxin or a radioisotope wherein said supplementary peptide sequence comprises a portion of an immunoglobulin.
 9. A soluble polypeptide fraction according to claim 8, wherein said supplementary peptide sequence comprises a portion of an immunoglobulin of IgG4 isotype.
 10. A method for producing the soluble polypeptide fraction of claim 6, which soluble polypeptide fraction further comprises a portion of an immunoglobulin, comprising the steps of:inserting a DNA molecule comprising a fusion of fragments of cDNA coding for the polypeptide regions corresponding to LAG-3 with cDNA coding for the portion of the immunoglobulin; transfecting the DNA molecule into a host expression system; and producing the soluble polypeptide fraction by expression in the host. 